Association of genetic polymorphisms of CISH with the risk of pulmonary tuberculosis in Zahedan, Southeast Iran

Abstract Background In the current study we aimed to find out the impact of cytokine-inducible Src homology 2 domain protein (CISH) gene polymorphisms on the risk of pulmonary tuberculosis (PTB) in a sample of Iranian population. Materials and methods Polymorphisms of CISH rs2239751, rs414171, and rs6768300 were determined in 200 PTB patients and 200 healthy subjects using T-ARMS-PCR or PCR-RFLP method. Results The results showed that rs414171 A>T genotypes significantly decreased the risk of PTB (OR = 0.16, 95% CI = 0.10–0.27, p < 0.0001, AT vs AA; OR = 0.31, 95% CI = 0.14–0.68, p < 0.0001, TT vs AA; OR = 0.19, 95% CI = 0.12–0.29, p < 0.0001, AT+TT vs AA; OR = 0.29, 95%CI = 0.20–0.42, p < 0.0001, T vs A). For rs6768300, the findings indicated that this variant decreased the risk of PTB (OR = 0.52, 95% CI = 0.33–0.82, p = 0.005, CG vs GG; OR = 0.57, 95% CI = 0.38–0.87, p = 0.012, C vs G). No significant association was observed between CISH rs2239751 polymorphism and risk/protection of PTB. Conclusion Our findings indicated that CISH rs414171 and rs6768300 variants might be associated with protection from PTB.

A clinicopathological study of primary central nervous system lymphomas & their association with Epstein-Barr virus.

Background & objectives: Primary central nervous system lymphomas (PCNSLs) are relatively uncommon, accounting for 2-3 per cent of primary brain tumours. Majority of these are diffuse large B cell lymphomas (DLBCL) occurring both in immunocompromised and immunocompetent patients. We undertook this study to classify PCNSL into germinal centre (GC) and non-germinal centre (NGC) type based on Hans classification and to find the role of Epstein-Barr virus (EBV) in pathogenesis both by conventional immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH). Methods: The consecutive cases of PCNSL during a 10 years period were analysed by IHC for CD45, CD20, CD3, B-cell lymphoma 2 and 6 (Bcl-2 and Bcl-6), B-cell specific octamer binding protein-1 (BOB-1), multiple myeloma oncogene-1 (MUM-1), EBV latent-membrane protein 1 (LMP-1), cyclin-D1, CD10, CD5 and CD23, as well as by CISH for EBV. Results: During a period of 10 years, 65 PCNSL were diagnosed which comprised 0.69 per cent (65/9476) of all intracranial tumours. The mean age of presentation was 49 yr with sex ratio (M:F) of 1.4:1. Most common location was supratentorial region with predominant involvement of frontal lobe. Single lesions were seen in 38 (58.4%) and multifocal lesions in 27 (41.5%) patients. None of the patients were immunocompromised. All cases were B cell immunophenotype and were DLBCL except one case of follicular lymphoma. According to Hans classification, majority of them were NGC (n=51, 79.6%) and 13 (20.3%) were GC type. Bcl-2 expression was noted in 34 (52.3%) tumours. EBV was positive in three (4.6%) cases; two were detected both by IHC and CISH and one case by CISH only. Interpretation & conclusions: In Indian population, PCNSL occurs mainly in immunocompetent patients, and a decade earlier than in western population. Immunophenotyping revealed that all cases were DLBCL with predominance of NGC type. No prognostic difference was seen between GC and NGC DLBCL. Association of EBV was rare and this virus was possibly not involved in the pathogenesis of PCNSL in immunocompetent individuals. CISH was an easy, economical and less cumbersome method for detection of EBV in PCNSL.

Evaluación de la amplificación del oncogén HER2 en pacientes con cáncer de mama a través de la técnica de hibridación in situ cromogénica (CISH) / Evaluation of HER2 oncogene amplification in breast cancer patients through technical chromogenic in situ hybridization

El protooncogén HER2 se encuentra localizado en el cromosoma 17. Codifica una glicoproteína transmembrana llamada receptor del factor de crecimiento epidérmico humano 2 (HER2). Está amplificado a bajos niveles en muchos tejidos normales y regula el crecimiento, diferenciación y muerte celular. La sobreexpresión de HER2 es el resultado de anormalidades en la amplificación del oncogén HER2. El objetivo de este trabajo fue evaluar la amplificación del oncogén HER2 en pacientes con cáncer de mama a través de la técnica de hibridación in situ cromogénica (CISH). La muestra estuvo constituida por 200 biopsias fijadas en formol al 10%, procesadas por las técnicas habituales hasta la inclusión en parafina, correspondientes a pacientes diagnosticadas con carcinoma ductal infiltrante de la mama, procedentes de consulta privada y del Instituto de Oncología "Dr. Miguel Pérez Carreño", con estudio inmunohistoquímico realizado en el Hospital Metropolitano del Norte en Valencia. Se encontró una asociación estadísticamente significativa (p<0,001) de la amplificación de HER2 con los diferentes niveles cuantitativos del producto de expresión del gen. Cabe destacar que hubo 16 (11%) de 146 casos considerados negativos desde el punto de vista inmunohistoquímico para la expresión de HER2, que resultaron amplificados para el oncogén a través de la CISH. Asimismo, más de la mitad (52,4%) de los casos positivos (3+) no presentaron amplificación. Este estudio demuestra la importancia de realizar la determinación de la amplificación de HER2 en los casos con resultado de 1+ por inmunohistoquímica, al evidenciar que el 12,3% de los casos son positivos a la amplificación del oncogén.

HER2 proto-oncogene is located on chromosome 17. It encodes a transmembrane glycoprotein called human epidermal growth factor receptor 2 (HER2). It is amplified at low levels in many normal tissues and regulates growth, differentiation and cell death. HER2 overexpression is the result of abnormalities in the HER2 oncogene amplification. The aim of this study was to evaluate the HER2 oncogene amplification in patients with breast cancer through the chromogenic in situ hybridization (CISH) technique. The sample consisted of 200 biopsies fixed in 10% formalin, processed by standard techniques to paraffin embedding, from patients diagnosed with infiltrating ductal carcinoma of the breast, coming from private practice and the Institute of Oncology "Dr. Miguel Pérez Carreño", with immunohistochemical study performed at Northern Metropolitan Hospital in Valencia. It was found a statistically significant association (p<0.001) of HER2 amplification with different quantitative levels of the product of the gene expression. It is noteworthy that there were 16 (11%) of 146 cases considered negative from the immunohistochemical point of view for the HER2 expression, that proved to be amplified for the oncogene through the CISH. Likewise, it is emphasized that more than half (52.4%) of the positive cases (3+) showed no amplification. This study shows the importance of the determination of HER2 amplification in cases result of 1+ by immunohistochemistry, demonstrating a 12.3% positive for these cases to the oncogene amplification.

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

Surgical complications in 1,049 cases of cervical intrafascial semm hysterectomy (CISH) / 대한산부인과학회지

OBJECTIVE: The aim of this study is to evaluate the complication rate of Cervical Intrafascial Semm Hysterectomy (CISH). METHODS: 1,049 CISH were carried out from April 1997 to August 2006 at our department. Retrospective review of medical records was performed to investigate the incidence and the type of complications during CISH procedures. RESULTS: 281 (26.79%) of the patients had a history of laparotomy. Overall 98 (9.34%) complications occurred. There were 30 (2.86%) transfusion, 19 (1.81%) transient bladder dysfunction, 12 (1.14%) cervical stump bleeding, 11 (1.05%) abdominal wound infection or disruption, 7 (0.67%) paralytic ileus, 6 (0.57%) incisional hernia, 4 (0.38%) cervical stump infection, 2 (0.19%) ureter injuries and 2 (0.19%) bladder injuries. And there were one case of transient peroneal nerve palsy, trocar site bleeding, trocar site endometriosis, foreign body in bladder and pelvic abscess. CONCLUSIONS: CISH procedure lessens the risk of ureteral and bladder injury, but this procedure increases the risk of cervical stump bleeding and infection morbidity.

Comparison of Her-2, EGFR and Cyclin D1 in Primary Breast Cancer and Paired Metastatic Lymph Nodes: An Immunohistochemical and Chromogenic In Situ Hybridization Study

The significant advance in the development of molecular-targeting drugs has made an evaluation of Her-2, EGFR, and cyclin D1 an important clinical issue in breast cancer patients. This study compared the Her-2, EGFR, and cyclin D1 status of primary tumors as well as their matching lymph node metastases using immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) in 73 breast cancer patients. Her-2, EGFR, and cyclin D1 protein showed a concordance between the primary lesion and the metastatic regional lymph nodes in 82%, 90%, and 63%, respectively. CISH also revealed 92%, 93%, and 85% concordance in the gene amplification status of Her-2, EGFR, and cyclin D1, showing a reasonable agreement between primary tumors and metastatic regional lymph nodes. Although a statistically significant agreement was found in Her-2 expression, a relatively high discordance rate (18%) raises a little concern. Our findings suggest that the Her-2 status can be reliably assessed on primary tumor but a possible difference can be found in Her-2, EGFR, and cyclin D1 status between the primary and the metastatic sites and this possibility should be concerned in patients considering molecular targeted therapy or patients with progress of disease.

Analysis of clinical factors influencing on the operation time of classical intrafascial supracervical hysterectomy (CISH) / 대한산부인과학회지

OBJECTIVE: The purpose of this study was to find clinical factors affecting the operation time of the classical intrafascial supracervical hysterectomy (CISH) performed on patients with uterine disease. METHODS: From January 2001 to December 2006, a total of 148 patients were reviewed and 130 patients with CISH were entered into the study. We investigated clinical factors, such as age, parity, BMI, height, weight, transverse diameter of uterus, previous operation number, operation assistant, period in sequence of operation date, pathology of uterus and operation time of surgery. The statistical assessment was used to find out the main factor determining the operation time of CISH. RESULTS: The mean operation time of CISH was 135.5 minutes and the mean transverse diameter of uterus was 7.2 cm. The main indications for CISH were uterine myoma or adenomyosis with dysmenorrhea, menorrhagia and vaginal bleeding. The operation time of CISH was affected by the transverse diameter of uterus and period which was categorized in sequence of operation date. CONCLUSION: Mainly, pre-operative ultrasonographic evaluation of uterine transverse diameter could predict the operation time of CISH.

Chromogenic in situ hybridization using routine tissue sections: MYCN in neuroblastoma.

Background: Amplification of the MYCN (myc myelocytomatosis viral related oncogene, neuroblastoma derived) gene in neuroblastoma is associated with a poor prognosis. Methods for estimating MYCN gene copy number that are based on pooled cells do not address copy number heterogeneity at the cell level and can underestimate or even miss amplification. MYCN copy number can be directly assessed by fluorescence in situ hybridization, but evaluation of tissue histology is difficult if not impossible. Objective: This paper reviews chromogenic in situ hybridization (CJSH) as it applies to the MYCN gene in neuroblastoma. We compare this technique to other methods for determining gene copy number and highlight the advantages of CISH. Methods: We have developed a chromogenic method for in situ hybridization (CISH) that enables us to determine MYCN copy number on an individual cell basis. This technique uses light microscopy on routine paraffin sections, and therefore allows simultaneous assessment of tumour histology. Results: In a previous study, CISH identified 100 % of the cases that were known to be amplified by other techniques and proved to be more sensitive than Southern blotting or the quantitative DNA polymerase chain reaction. The MYCN copy number is generally believed not to vary within a tumour, nor between tumour samples, including primary vs. metastases, and pre-and post-treatment specimens. However, we found heterogeneity from cell to cell, with ~30 % of amplified tumours showing >50 % variation in MYCN copy between cells. Conclusion: For detection of gene amplification, CISH has all the advantages of FISH but in addition, needs no special microscopy or image capturing systems, and preparations are permanent. In the case of neuroblastoma, CISH has disclosed considerable heterogeneity in MYCN copy number between cells in a tumour. Heterogeneity reflects different tumour clones and its role has been under-recognized in neuroblastoma biology. Additional studies are needed to investigate the significance of tumour heterogeneity in neuroblastoma, and whether the aggressive (i.e., MYCN-amplified) clones are more likely to metastasize, survive treatment modalities, and ultimately kill the patient.

Amplification of the HER-2/neu Oncogene and Topoisomerase II-alpha by Chromogenic in Situ Hybridization in Breast Cancer

PURPOSE: HER-2/neu is the most frequently amplified oncogene in breast cancer. Topoisomerase II-alpha is a key enzyme in DNA replication and it is a molecular target for many anti-cancer drugs that are called topo II inhibitors; in addition, it is a new marker of proliferation. Because of the physical proximity of the ER-2/neu and topoisomerase II-alpha genes, co-amplification of the HER-2/neu and topoisomerase II-alpha may be important determinates of the response to chemotherapy for advanced breast cancer patients. METHODS: We studied the correlation of gene amplification of HER-2/neu and topoisomerase II-alpha by chromogenic in situ hybridization (CISH) in 43 infiltrating duct carcinomas of the breast. The over-expression of HER-2/neu protein and the staining index for the proliferation marker of topoisomerase II-alpha were examined immunohistochemically. The correlations between the status of HER-2/neu and topoisomerase II-alpha and the other clinicopathologic variables such as tumor size, lymph node metastasis, TNM stage, histologic grade, nuclear grade, and the estrogen receptor and progesteron receptor were investigated. RESULTS: Of the 43 infiltrating ductal carcinomas, the amplifications of HER-2/neu and topoisomerase II-alpha by CISH were observed in 8 cases (18.6%) and 14 cases (32.6%), respectively. Amplification of HER-2/neu showed the statistically significant correlations with tumor size, histologic grade and the topoisomerase II-alpha staining index. Amplification of topoisomerase II-alpha showed statistically significant correlations with axillary lymph node metastasis, the stage, the nuclear grade and the estrogen receptor status. CONCLUSION: These data suggest that amplification of HER-2/neu oncogene and topoisomerase II-alpha by CISH may be valuable for determining the response to chemotherapy, and detection of HER-2/neu and topoisomerase II-alpha in tumor sections may have prognostic value in human breast cancer.

The Clinical Results of Classic Intrafascial Supracervical Hysterectomy for 10 Years / 대한산부인과학회지

OBJECTIVE: We have performed Classic Intrafascial Supracervical Hysterectomy (CISH) since April 1993, so we are to report the clinical results of CISH for 10 years, and introduce the technical aspects of new CISH. METHODS: A retrospective analysis on 470 cases of CISH (conventioal and new method) was carried out including age, parity, operating time, blood loss, uterine weight, pathologic results, complications and postoperative recovery. RESULTS: The mean age was 44.8 years (range 25-68), parity 2.23 (0-9), operating time 131 minutes (70-310), mean blood loss 148 mL (20-1000) and mean uterine weight was 306 gram (90-880). The most common uterine pathology was uterine leiomyoma (49.3%) and there were adenomyosis (15.5%), leiomyoma with adenomyosis (12.5%), chronic cervicitis (5%) and so on. Intraoperative complications included one case of bladder injury due to thick adhesion and 6 cases of bleeding requiring transfusion. Postoperative complications included 5 cases of cervical coring site bleeding, 2 cases of febrile complications, 2 cases of trocar site bleeding, 1 case of transient nerve injury of lower extremity, and 1 case of urinary tract infection. The mean hospital stay was 5.3 days (3-10). two hundreds and seventy eight patients of 470 had been followed for cervical pathology, and no pathologic findings except 3 cases of mild dysplasia were shown. CONCLUSION: Classic intrafascial supracervical hysterectomy, a minimally invasive organ-preserving procedure indicated for benign uterine disease, is associated with few operative complications and low morbidity, and the disadvantages of conventional Semm's CISH technique can be overcome by new CISH technique using triple ligation method.

Modified Classic Intrafascial Supracervical Hysterectomy: The comparison with conventional method / 대한산부인과학회잡지

OBJECTIVE: To introduce the technical aspects and advantages of a new classic intrafascial supracervical hysterectomy (CISH) technique over the conventional technique. METHODS: The charts of two hundred women who underwent conventional CISH technique (100 cases) between March 2000 and September 2000, or the new CISH technique (100 cases) between May 2002 and November 2002 were reviewed regarding patient characteristics, indications, uterine weight, estimated blood loss, operating time, and hemoglobin change. Cases with a very large uterus (over 500 g of weight), severe adhesions, and other complications, which required a longer operating time, were excluded and this study compared the most recent patient's data for both procedures, since the surgeon's experience and surgical technique will improve with practice. RESULTS: The women who underwent the new CISH had significantly shorter operating time as compared with the conventional method (106.7 vs 90.4 min). Although there was no significant difference in the estimated blood loss, the drop of Hb, which is an objective sign of blood loss, was significantly smaller using the new CISH technique than using the conventional CISH technique (1.98 vs 1.64 g/dL). CONCLUSION: The new CISH technique is safer, more convenient with less blood loss and shorter op time than the conventional technique, especially when the uterus is markedly enlarged by a large myoma, the ovarian ligament is too short, or the ovary and uterus are very closely adherent.

Chromogenic In Situ Hybridization Analysis to Determinate HER-2/neu Status in Breast Carcinoma

PURPOSE: The determination of HER-2/neu gene amplification has become necessary for the selection of breast cancer patients to undergo anti-HER-2/neu therapy, using a humanized monoclonal antibody. Chromogenic in situ hybridization (CISH) detection of the HER-2/neu gene, a newly developed method, utilizes a robust and unique-sequence DNA probe labeled with digoxygenin, which is sequentially incubated with antidigoxygenin fluorescein, antifluorescein peroxidase and diaminobenzidine. The aim of this study was to establish a CISH assay for the detection of HER-2/neu amplification. The results were compared with those of the immunohistochemistry (IHC) methods, most frequently used for detecting HER-2/neu alteration. METHODS: CISH was performed in 4 groups of infiltrating breast carcinomas. Each group was comprised of 20 cases in which the HER-2/neu stati had previously been scored on a four value scale: 0, 1+, 2+ and 3+ by IHC. The results of CISH and IHC were compared for each tumor group. The HER-2/neu gene amplification detected by CISH was thpically visualized as large DAB-stained clusters or by many dots in the nucleus. RESULTS: The concordance between the CISH and IHC was 95% (kappa=0.901). Three IHC-positive cases (score 2+) showed no gene amplification and one IHC-negative case (score 1+) showed gene amplification by CISH. CONCLUSION: The current study showed excellent agreement between the CISH and IHC methods. CISH is an accurate, practical and economical approach for determining the HER-2/neu stati in breast carcinomas. It is also a useful methodology for confirming the IHC results in paraffin- embedded tumor samples, so offers a promising alternative to IHC in a routine diagnostic setting.

The clinical efficacy of pelviscopic classic intrafascial supracervical hysterectomy / 대한산부인과학회잡지

OBJECTIVES: Our purpose was to evaluate the efficacy of pelviscopic classic intrafascial supracervical hysterectomy (CISH) & compare it with total abdominal hysterectomy (TAH). METHODS: We analyzed retrospectively, the clinical data of 1126 patients admitted from January 1993 to December 1998, at 11 university or general hospitals in Korea, for CISH group and 363 patients admitted from January 1993 to December 1998, at Chung-Ang university hospitals, Seoul, Korea, for TAH group. These patients were operated on with the indication of benign uterine disease without cervical malignant lesion. The patients with malignant cervical lesion were excluded in this study. RESULTS: (1) The average age of CISH group was 42.2 (range 24-63) years old and TAH group was 46 (range 31-54) years old. (2) The mean operative time was 150.7+/-49.9 (mean+/-SD, range 55-395) minutes in CISH group and 133.8+/-35 (mean+/-SD, range 65-350) minutes in TAH group. The mean estimated blood loss was 206+/-183.6 (mean+/-SD, range 20-2000) ml in CISH group and 596+/-452.3 (mean+/-SD, range 100-6500) ml in TAH group. The mean hemoglobin change was 1.52+/-0.98 (mean+/-SD, range 0-7) g/dL in CISH group and 3.03+/-1.47 (mean+/-SD, range 0.2-8.4) g/dL in TAH group. The mean time in hospital was 6.3+/-1.43 (mean+/-SD, range 3-18) days in CISH group and 11.0+/-4.08 (mean+/-SD, range 7-37) days in TAH group. (3) Overall complications occured at 5.3% in CISH group and 9.1% in TAH group. (4) Pathologic findings of cervix were benign in 1121 patients (99.6%) and severe dysplasia or carcinoma in situ in 5 patients (0.4%) among CISH groups. In TAH groups, 351 patients (96.7%) had benign cervical lesion and 12 patients (3.3%) had more than oderate dysplasia or carcinoma in situ. In both groups, no invasive cervical cancer was found and the transformation zone of the cervix was included within the resection margin of the cervix in all cases. CONCLUSION: The pelviscopic CISH group has less operative blood loss, less postoperative hemoglobin change, shorter hospital stay and less operative complication than TAH group. Pelviscopic CISH is truly a minimally invasive and organ-preserving surgery and coring out the cervix with the calibrated uterine resection tool may prevent the development of cervical cancer. Therefore, we believe that pelviscopic CISH is preferred in cases of benign uterine diseases, because it lowers operative complications and it may have several benefits compared with total hysterectomy, especially, in women with no pathologic lesion of the cervix.

Clinical Evaluations of the Classical Intrafascial SEMM Hysterectomy (CISH) / 대한산부인과학회잡지

Objectives: We performed this study to review the clinical availability of the classical intrafascial SEMM hysterectomy (CISH), and report our experience for 2 years restrospectively. METHODS: From March 1999 to February 2001, 100 cases of CISH were performed at the Department of Obstetrics and Gynecology, College of Medicine, Kangdong Sacred Heart Hospital, Hallym university. We performed this procedure by 3 steps. The dissection of uterus was done by classical suture methods (including extracorporeal and intracorporeal suture techniques) to uterine isthmic portion. And then transvaginal cylindrical coring out of the cervical tissue was done using a cervix corer, the Calibrated Uterine Resection Tools (CURT) for resection of cervical tissue especially transformation zone. Finally, the dissected uterus was extracted by using a Serrated Edged Macro-Morcellator (SEMM) without colpotomy. We reviewed the patient's data retrospectively and made phone calls to each patients. RESULTS: The indications for CISH, based on the pre-and intraoperative diagnosis, were leiomyoma, adenomyosis, endometrial hyperplasia and chronic pelvic inflammatory disease or pelvic pain, in order of frequency. The mean operative time was 126 minutes and the mean hemoglobin change was 1.27 gm/dL. The mean hospital days were 5.7 days and the mean age of the patients was 44.6 years old. There were no definite intra- and postoperative complications except for 3 cases of CO2 retentions and 2 cases of delayed bleeding on cervical stump. The mean days to first sexual contact after surgery were 34.5 days. In view of sexual feeling, 86% of the patients didn't feel any difference and only 13% felt worse than before. During postoperative follow-up, the cervical cytologic study were found no endocervical cells at all of 44 checked patients. CONCLUSION: The CISH procedures can be safely performed by well-trained laparoscopists, and this technique minimally alters the anatomy and integrity of the pelvic floor and vagina. Therefore the reductions of surgical morbidity, blood loss, hospitalization and postoperatve discomfort were obtained. Through the preservation of the anatomical relations of the pelvic floor and the function of female sexuality, this technique is the new proposal in gynecologic surgery.

Chromosomal instability in oral premalignancy / 대한내과학회지

BACKGROUND: Squamous cell carcinoma of the oral cavity was thought to develop through a multistep process driven by genetic changes. Recently, the use of in situ hybridization on paraffin sections has been reported, thus allowing one to visualize specific chromosomal changes without disrupting the spatial distributions of such changes in the tissue samples. Specific aims of this study were to determine whether genetic alterations are present in premalignant oral lesions, i.e. leukoplakia and to examine whether the degree of genetic alterations is associated with risk of cancer development. METHODS: Twenty-eight formalin fixed, paraffin- embedded specimens were obtained from 26 patients with oral leukoplakia at the University of Texas MD Anderson Cancer Center. The 6-micron thick tissue sections were mounted on glass slides precoated with silane or poly-l- lysine. The tissue sections were baked at 65oC overnight and deparaffinized in xylene-ethanol series. And then presoaked in cold pepsin/HCl solution for 15 minutes and incubated with 0.4% pepsin in 0.2N HCl at 37oC for 10-45 minutes. The biotinylated centromeric DNA probe specific for chromosome 17 and target DNA were denatured together at 90oC-95oC for 4-7 minutes and incubated at 37oC overnight in sealed wet chamber. The tissue sections were stained with immunoperoxidase technique using avidin-biotin-peroxidase enzyme complex and diaminobenzidine as a substrate. The tissue sections were counterstained with Giemsa stain. At least, 200 nuclei were scored in each slide. The total number of signal spots were divided by the number of nuclei analyzed to obtain a chromosome index. The cells with > or = 3 chromosome copies were divided by the total counting cells to obtain the degree of chromosomal polysomy. RESULTS: Chromosomal polysomy was more prevalent in dysplastic lesions (median 8.1%, range 0.5-23.0%) than lesions without dysplasia (median 3.0%, range 0-14.5%). Interestingly, 6/17 cases exhibiting >2.0% polysomy subsequently developed invasive carcinoma or carcinoma in situ compared to 0/11 cases with low or no polysomy. There was clear association between chromosome index and the degree of chromosomal polysomy. CONCLUSON: Oral premalignant lesions had genetic alterations. Dysplastic lesions habor more genetic changes than hyperplastic lesions. Risk of oral cancer development seems to be associated with cummulative genetic changes. There was clear association between chromosome index and the degree of baseline chromosomal polysomy. These results suggest that the degree of chromosomal instability present in premalignant oral lesions is associated with the risk of cancer development.

CISH ( Classic Intrafascial SEMM Hysterectomy ) for Huge Uterine Leiomyomata / 대한산부인과학회잡지

Since hysterectomy using laparoscopy has been first reported in 1989, various techniques for laparoscopic hysterectomy were developed. However, complications during hysterectomy still remained and many clinicians assert that huge uterine masses should be performed with laparotomy for safety. We have performed CISH for benign uterine disease such as uterine leiomyomata, PID, chronic pelvic pain, DUB and adenomyosis, etc since 1993. This report is to evalute the clinical efficacy and the feasibility of the CISH for huge uterine masses, which include 20 patients who underwent CISH with huge uterine leiomyomata(more than 250 gm and /or 14 gestational size) between June 1995 and May 1996 in Chung-Ang University Pil-Dong Hospital. No laser, monopolar and stapling cutting devices were used. All of the hysterectomy procedures were performed in the classic manner with grasping forceps, scissors, ligatures, and sutures. We excluded the patients who had cervical neoplasia, or who wanted to have total removal of the uterus in this study. The results were as follows. 1) The age of patients were ranged from 37 to 50 years, and the mean age was 43.78+/-3.8 years. 2) The parity distribution of patients were ranged from 0 to 4, the mean was 1.93+/-0.9. 3) The mean operative time was 171.78+/-36.5minutes, ranged from 130 to 240 minutes. 4) The mean estimated blood loss(EBL) was 223.8+/-206.7cc, ranged from 40 to 700cc 5) The mean uterine weight was 345.9+/-87.3gm, ranged from 256 to 625.7gm 6) There was no major complications and a few minor complication. No procedure was converted to laparotomy. This report suggests that the CISH technique for benign huge uterine leiomyomata is feasible, safer technique, and associated with significantly low morbidity and no major complication.