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흑염소의 체내수정란 생산에 관한 연구

  • 최창용;조숙현;한만희;권응기;최성복;최연호;최순호;손동수;최상용
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2002년도 국제심포지엄
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    • pp.86-86
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    • 2002
  • 면양과 염소가 최근 수십년동안 세계여러 나라에서 번식생리의 연구를 위한 모델로 사용되어 왔는데, 체내수정란의 생산에 관한 영역도 유럽을 중심으로 활발하게 연구되어왔다. 수정란생산을 위한 발정동기화방법, 과배란처리 및 수정란회수방법 기술은 현재 상당히 많은 기술진척이 이루어진 상태이나, 우리나라 고유의 재래유전자원인 흑염소에는 이를 위한 기술이 미진한 실정이므로 본 실험에서는 흑염소의 체내수정란생산기술을 확립하여 재래가축 유전자원보존을 위한 기초기술을 마련하고자 한다. 축산기술연구소 남원지소에서 사육하고 있는 체중 20kg 이상의 건강한 흑염소를 이용하여 발정동기화를 위해 controlled intravaginal drug release(CIDR)를 질내에 14일 동안 삽입하고, 과배란처리는 FSH를 CIDR 삽입 12, 13, 14일째에 12시간 간격으로 점감법으로 총20mg을 투여하였으며, PGF$_2$a를 13일째 FSH와 함께 투여하였다. CIDR는 14일째의 아침에 제거하였다. 수컷과의 교미는 CIDR제거 24시간후에 GnRH를 투여와 동시에 실시하였으며, 채란은 교미후 3일째에 외과적인 방법으로 실시하였다. CIDR처리경과에 따른 progesterone농도는 CIDR 주입시 바로 수치가 상승하여 제거전까지 6~12ng/m1의 농도를 유지하였으며, 제거즉시 2ng/ml 이하로 떨어졌다. 채란시 평균 배란점은 16.5개, 미배란난포 9.8개였으며, 회수수정란은 6.0개를 나타내어 채란율은 36.4%를 나타내었다. 회수된 수정란의 발달단계는 4-cell 78.9%, 2-cell 5.3%, fragmentation 15.8%를 나타내었다. 이와 같은 체내수정란생산방법을 기반으로 하여 이후 수정란의 동결 및 수정란이식기법에 관한 연구를 수행한다면 우리나라의 재래가축인 흑염소의 유전자원 장기보존과 생산성향상에 기여할 것으로 사료된다.배양액에 30 embryos/50ul 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$의 탄산가스 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT를 이용하여 통계분석을 실시하였다. 체외수정 12시간 후에 난자 급속 염색법으로 염색을 실시한 결과, 모든 처리구에서 핵성숙률(76.4~95.2%), 정자침투율(51.1~66.9%), 웅성전핵형성률(95.2~100%), 다정자침입률(18.2~25.6%) 및 평균침입정자수(1.2~l.4개)에서 유의적인 차이가 인정되지 않았다. 체외배양 48시간 난할률을 조사한 결과, 처리구별 차이(53.9~67.9%)는 인정되지 않았으나, 배양 7일째 배반포형성률은 각각 14.5, 25.4, 17.3 및 12.4%로서 25uM의 $\beta$-ME처리구가 유의적(P<0.05)으로 높은 배발달률을 나타내었고, 총세포수에 있어서는 대조구와 처리구간 유의적인 차이가 인정되지 않았다. 따라서 돼지 난포란을 성숙배양할 때, 25uM $\beta$-ME를 첨가배양하는 것이 양질의 돼지체외수정란을 생산하는 하나의 방법으로 조사되었다.다.natural objects and was popular at the time of Yukjo Dynasty, and there are some documents of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to have been "Japanese Letter Jobcheso." Therefore, the purpose of this study is to look into the origin

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분만 직전의 개에서 급성 부신겉질저하증과 유사한 일과성 저나트륨혈증 및 고칼륨혈증 일례 (Hypoadrenocortical Crisis-like Transient Hyponatremia and Hyperkalemia in a Near-Term Pregnant Dog)

  • 강지훈;김민준;조민행;장동우;강현구;김일화;나기정;양만표
    • 한국임상수의학회지
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    • 제22권4호
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    • pp.450-452
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    • 2005
  • A 3-year-old, 43kg, pregnant Great Pyrenese was examined for clinical signs of acute weakness and anorexia for 4 days. The dog was in lateral recumbency at referral. The rectal temperature was within reference range, and the respiratory and heart rates were 36 breaths/min and 58 beats/min, respectively. The abdomen was distended, and several puppies were palpated. The mean fetal head diameter was 2.8cm in the ultrasonographic examination. The initial complete blood count and serum biochemical examinations revealed mild dehydration, mud hyperglycemia, hypochloremia, hyperkalemia, hyponatremia, and low sodium-potassium ratio. Serum BUN and total cholesterol values were slightly high. Hypoadrenocortical crisis was suspected on the basis of signs of acute collapse, hyponatremia and hyperkalemia. Adrenal gland function was evaluated by an ACTH stimulation test. The baseline cortisol concentration was $18.6{\mu}g/dl$ and the concentration at 1 hour after administration of tetracosactrin (ACTH, Synacthen) was $8{\mu}g/dl$. The dog was treated for the correction of assumed hypoadrenocortical crisis and substantial hyperkalemia. In addition to rapid infusion with saline solution, other medications administered intravenously included sodium bicarbonate and cimetidine hydrochloride. The dog was monitored with repeated serum electrolyte examination. After clinical stabilization, cesarean section was performed. All of 13 puppies were delivered, and the dog recovered from anesthesia without complications. The values of postpartum blood tests returned to normal or within reference range. The dog remained healthily.

Urinary Estrone Sulfate for Monitoring Pregnancy of Dairy Cows

  • Yang, C.J.;Wu, L.S.;Tseng, C.M.;Chao, M.J.;Chen, P.C.;Lin, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권9호
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    • pp.1254-1260
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    • 2003
  • The purpose of this study firstly was conducted to establish a radioimmunoassay (RIA) of estrone sulfate ($E_1S$), secondly to monitor the reproductive status of dairy cows using their urine samples. Urine and blood samples were collected in series within a day from four pregnant Holstein-friesian cows to evaluate the relationship between $E_1S$ levels in blood and urine with or without urinary creatinine basis. The urine was then collected biweekly from three cows in estrous and those artificially inseminated; collection from pregnant cows was made on a monthly basis. Results indicated that sensitivity for the $E_1S$ RIA was 5 pg/tube and the recovery rate was 100%. The daily urinary creatinine concentrations fluctuated within a day, but changes were slighter in midday, whereas the changes of concentrations of $E_1S$ in urine were relatively smaller. The concentrations of serum $E_1S$ during the estrous cycle were undetectable due to the limitation of assay, but the urinary $E_1S$ level could be measured with no obvious changes during the cycle. The urinary $E_1S$ levels increased remarkably around 7.7 to 8.3 ng/ml, 80 to 100 days after pregnancy but the serum $E_1S$ levels did not elevate until 120 to 150 days. The level of $E_1S$ increased gradually during pregnancy and eventually reached its peak before parturition at around 40 ng/ml and finally decreased to its basal level 2 days postparturition. During pregnancy, $E_1S$ concentrations of urine increased earlier than those in blood. The correlation coefficients between urinary and serum $E_1S$ concentration during pregnancy and postparturm were higher than those adjusted with creatinine (creatinine ratio). The concentrations of $E_1S$ in urine could be maintained unchanged for 8 days storing the samples in room temperature, which was extended to 8 days when the samples were pretreated by boiling for 30 minutes or treated with autoclave. In conclusion urinary $E_1S$ concentrations can be used directly for monitoring the pregnant status and fetal viability of dairy cows and can assist accurate confirmation of pregnancy in cows at least 80 to 100 days after insemination much earlier than by serum $E_1S$.

IN VITRO DEVELOPMENT OF BOVINE ONE-CELL EMBRYOS FERTILIZED IN VITRO IN SERUM- AND FEEDER CELL-FREE CULTURE SYSTEMS

  • Ohboshi, S.;Hanada, K.;Zhao, J.;Hattori, M.;Fujihara, N.;Umetsu, R.;Yoshida, T.;Tomogane, H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제9권5호
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    • pp.583-590
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    • 1996
  • The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium (mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera (CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage (8.6-21.4%), dependent upon their product lots, although the development might not be affected at least by serum progesterone levels. ${\beta}$-Mercaptoethanol (${\beta}$-ME) supplemented into mSOFM was effective to the embryonic development (27.8%), as well as the co-culture system with cumulus cells (19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and ${\beta}$-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin (BSA, fraction V), but not polyvinyl alcohol (PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture (CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA (7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium (final 2.0 mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with ${\beta}$-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos.

Relationship between erb-B2 mRNA Expression in Blood and Tissue of Invasive Ductal Carcinoma Breast Cancer Patients and Clinicopathological Characteristics of the Tumors

  • Moazzezy, Neda;Ebrahimi, Fatemeh;Sisakht, Mahsa Mollapour;Yahyazadeh, Hossein;Bouzari, Saeid;Oloomi, Mana
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권1호
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    • pp.249-254
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    • 2016
  • Molecular detection methods such as RT-PCR for detecting breast cancer-associated gene expression in the peripheral blood have the potential to modify breast cancer (BC) staging and therapy. In this regard, we evaluated the potential of erb-B2 molecular marker in BC detection and analyzed the expression of erb-B2 mRNA in the peripheral blood and fresh tissue samples of 50 pretreated female BC patients and 50 healthy females by reverse transcription-PCR (RT-PCR) method. We also assessed the correlation of erb-B2 mRNA marker positivity in peripheral blood and tumor tissue samples with clinical and pathological factors in BC patients in order to evaluate its prognostic value. It was shown that there is a significant difference between healthy females and BC patients with expression of the erb-B2 molecular marker (p<0.01). A significant difference between the expression of erb-B2 in the peripheral blood and tissue samples of BC patients (p<0.01) and the frequency of circulating erb-B2 mRNA expression in peripheral blood and in tissue was detected by RT-PCR. No correlation was found between erb-B2 mRNA expression in blood or tumor tissue samples and lymph node, tumor grade, tumor stage, tumor size, patient's age, ki67, estrogen receptor (ER), progesterone receptor (PGR), P53, and HER-2 status. However, in a small subset of 31 BC patients we found that expression of erb-B2 in peripheral blood or in both peripheral blood and tumor tissue was directly correlated with lympho-vascular invasion and perineural invasion as poor prognostic features. The highest rates of erb-B2 expression in peripheral blood or tumor tissue were in the ER and PR negative and HER-2 positive group. This study suggests that the application of the RT-PCR and immunohistochemical methods for erb-B2 molecular marker detection would provide a higher detection rate, especially in early stage BC.

Drug Use Evaluation of Letrozole in Breast Cancer Patients at Regional Cancer Hospitals in Thailand

  • Ketkaew, Chaninun;Kiatying-Angsulee, Niyada
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권14호
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    • pp.6055-6059
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    • 2015
  • Background: Medication policy development in Thailand is continually promoting rational drug use. Letrozole, an endocrine therapy drug, is usually prescribed for post-menopausal status early and advanced stage breast cancer. After Ministry of Public Health announced Letrozole as compulsory licensed drug in 2009, more breast cancer patients can access to this drug at low cost especially those within universal coverage schemes. To ensure that Letrozole is rationally prescribed, the drug utilization study was conducted. Objectives: The aim of this study was to describe the appropriate use of Letrozole in breast cancer and the relationship between appropriate use and health benefit schemes. Materials and Methods: A retrospective study to evaluate use of Letrozole in breast cancer patients was performed for six months between January - June 2010 in seven regional cancer hospitals, Thailand. All prescriptions of Letrozole were identified from pharmacy dispensing databases and prescription papers. A medical record review was also performed to evaluate appropriate use referring to the drug use evaluation criteria. The approved criterion of this study was referred from the guideline of Thai National Formulary version 2010. Results: There were 681 prescriptions of Letrozole for 254 breast cancer patients with an average age of $58.6{\pm}10.0years$. The patients in universal coverage scheme (UCS), civil servant medication benefit scheme (CSMBS) and social security scheme (SSS) were 77.7%, 18.5% and 8.7% respectively. 10.6% were prescribed Letrozole for the first time. Letrozole were prescribed by oncologists (82.8%). The average number of tablets per prescription was $58{\pm}10$. Calcium supplements were prescribed concomitant with Letrozole for 19.4%. To assess drug use evaluation criteria, 45 prescriptions were excluded because of uncompleted clinical data, 636 prescriptions were evaluated. The study showed 86 prescriptions (13.5%) with inappropriate use including 6 (0.9%) not prescribed for estrogen receptor (ER) and/or progesterone receptor (PR) positive, 31 (4.9%) not prescribed for post-menopausal and 49 (7.7%) not prescribed for an appropriate duration. Appropriate use percentages in different health benefit schemes were similar, 85.7% of CSMBS, 86.4% of SSS and 86.7% of UCS. The relationship between health benefit scheme and appropriate use of Letrozole was not significantly different, ${\chi}^2$ (2, N = 636) = 0.081, p > 0.05. Conclusions: The study showed inappropriate use in breast cancer patients because of non-compliance with duration, menopausal status and hormone receptor requirements. To prescribe appropriate indication did not referred to the appropriate practice along the treatment. Drug use evaluation proved very useful for detecting the sign of inappropriate use and allows immediate feedback to the stakeholder for developing medication policy in the future. Importantly, there was no significantly difference in appropriate use of Letrozole across health benefit schemes.

Immunohistochemical Profile of Breast Cancer Patients at a Tertiary Care Hospital in New Delhi, India

  • Doval, Dinesh Chandra;Sharma, Anila;Sinha, Rupal;Kumar, Kapil;Dewan, Ajay Kumar;Chaturvedi, Harit;Batra, Ullas;Talwar, Vineet;Gupta, Sunil Kumar;Singh, Shailendra;Bhole, Vidula;Mehta, Anurag
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권12호
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    • pp.4959-4964
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    • 2015
  • Background: To assess the immunohistochemical expression of estrogen receptor (ER), progesterone receptor (PgR) and human epidermal growth factor receptor-2 (HER2) neu receptor in breast cancer and their associations with various clinicopathological characteristics. Materials and Methods: This is a retrospective analysis of women who presented with primary, unilateral breast cancer in the Department of Medical Oncology at Rajiv Gandhi Cancer Institute and Research Centre, Delhi, India during the period from January 2008 to December 2011. Data were retrieved from the medical records of the hospital including both early and locally advanced cancer cases. ER, PgR and HER2neu expression in these patients was assessed and triple negative patients were identified. Associations of triple negative and non-triple negative groups with clinicopathological characteristics were also evaluated. Results: A total of 1,284 women (mean age 52.1 years, 41.9% premenopausal) were included in the analysis. Hormone receptor positivity (ER and/or PgR) was seen in 63.4% patients, while 23.8% of tumors were triple negative. Only 23.0% were HER2 positive. Around 10.0% of tumors were both ER and HER2 positive. ER and PgR positivity was significantly associated with negative HER2 status (p-value <0.0001). Younger age, premenopausal status, higher tumor grade, lymph node negativity, advanced cancer stage, and type of tumor were strongly associated with triple negativity. Significantly, a smaller proportion of women had ductal carcinoma in situ in the triple negative group compared with the non-triple negative group (35.6% versus 60.8%, p-value<0.01). Conclusions: The present analysis is one of the largest studies from India. The majority of the Indian breast cancer patients seen in our hospital present with ER and PgR positive tumors. The triple negative patients tended to be younger, premenopausal, and were associated with higher tumor grades, negative lymph nodes status and lower frequency of ductal carcinoma in situ.

Multivariate Analysis of Prognostic Factors in Male Breast Cancer in Serbia

  • Sipetic-Grujicic, Sandra Branko;Murtezani, Zafir Hajdar;Neskovic-Konstatinovic, Zora Borivoje;Marinkovic, Jelena Milutin;Kovcin, Vladimir Nikola;Andric, Zoran Gojko;Kostic, Sanja Vladeta;Ratkov, Isidora Stojan;Maksimovic, Jadranka Milutin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권7호
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    • pp.3233-3238
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    • 2014
  • Background: The aim of this study was to analyze the demographic and clinical characteristics of male breast cancer patients in Serbia, and furthermore to determine overall survival and predictive factors for prognosis. Materials and Methods: In the period of 1996-2006 histopathological diagnosis of breast cancer was made in 84 males at the Institute for Oncology and Radiology of Serbia. For statistical analyses the Kaplan-Meier method, long-rank test and Cox proportional hazards regression model were used. Results: The mean age at diagnosis with breast cancer was $64.3{\pm}10.5$ years with a range from 35-84 years. Nearly 80% of the tumors showed ductal histology. About 44% had early tumor stages (I and II) whereas 46.4% and 9.5% of the male exhibitied stages III and IV, respectively. Only 7.1% of male patients were grade one. One-fifth of all patients had tumors measuring ${\leq}2cm$, and 14.3% larger than 5 cm. Lymph node metastasis was recorded in 40.4% patients and 47% relapse. Estrogen and progesterone receptor expression was positive in 66.7% and 58.3%, respectively. Among 14.3% of individuals tumor was HER2 positive. About two-thirds of all male patients had radical mastectomy (66.7%). Adjuvant hormonal (tamoxifene), systematic chemotherapy (CMF or FAC) and adjuvant radiotherapy were given to 59.5%, 35.7% and 29.8% patients respectively. Overall survival rates at five and ten years for male breast cancer were 55.0% and 43.9%, respectively. According to the multivariate Cox regression predictive model, a lower initial disease stage, a lower tumor grade, application of adjuvant hormone therapy and no relapse occurrence were significant independent predictors for good overall survival. Conclusions: Results of the treatment would be better if disease is discovered earlier and therefore health education and screening are an imperative in solving this problem.

Inflammatory Breast Cancer: a Single Centre Analysis

  • Gogia, Ajay;Raina, Vinod;Deo, Suryanarayan Vishnu;Shukla, Nootan Kumar;Mohanti, Bidhu Kalyan;Sharma, Daya Nand
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권7호
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    • pp.3207-3210
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    • 2014
  • Background: Inflammatory breast cancer (IBC) is an aggressive form of locally advanced breast cancer characterized by rapidly progressive breast erythema, pain and tenderness, oedema and paeu d'orange appearance. It accounts for 1-3% of all newly diagnosed cases of breast cancer in the west. Data on IBC from India are lacking. The aim of our study was to assess the clinical-pathological parameters and outcome of IBC at, All India Institute of Medical Sciences, a large tertiary care centre. Materials and Methods: We screened 3,650 breast cancer cases registered from January 2004 to December 2012 and found 41 cases of IBC. Data included demographics as well as clinical, radiological and histopathological characteristics, and were collected from clinical case records using the International Classification of Diseases code (C-50). Patients who presented with IBC as a recurrence, or who had a neglected and advanced breast cancer that simulated an IBC were excluded from this study. Results: The median age was 45 years (range 23-66). The median duration of symptoms was 5 months. The American Joint Committee on Cancer stage (AJCC) distribution was Stage III - 26 and IV - 15 patients. Estrogen receptor (ER), progesterone receptor (PR) positivity and human epidermal growth factor receptor 2 (HER2/neu) positivity were 50%, 46% and 60%, respectively. Triple negativity was found in 15% of the cases. All the non metastatic IBC patients received anthracycline and/ or taxane based chemotherapy followed by modified radical mastectomy, radiotherapy and hormonal therapy as indicated. Pathological complete remission rate was 15%. At a median follow-up of 30 months, the 3 year relapse free survival and overall survival were 30% and 40%respectively. Conclusion: IBC constituted 1.1% of all breast cancer patients at our centre. One third of these had metastatic disease at presentation. Hormone positivity and Her2 neu positivity were found in 50% and 60% of the cases, respectively.

Clinico-Pathologic Subtypes of Breast Cancer Primary Tumors Are Related to Prognosis after Recurrence

  • Sanchez, Cesar;Camus, Mauricio;Medina, Lidia;Oddo, David;Artigas, Rocio;Sepulveda, Alejandra Perez;Domainguez, Francisco;Razmilic, Dravna;Navarro, Maria Elena;Galindo, Hector;Acevedo, Francisco
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권12호
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    • pp.5081-5086
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    • 2016
  • Background: Pathological factors, based mainly on immunohistochemistry (IHC) and histological differentiation, are mostly used to differentiate breast cancer (BC) subtypes. Our present aim was to describe the characteristics and survival of a relapsing BC patient cohort based on clinico-pathologic subtypes determined for the primary tumors. Methods: We used a clinico- pathological definition of BC subtypes based on histological grade (HG), estrogen receptor (ER), progesterone receptor (PgR),and epidermal growth factor receptor type 2 (HER2) expression assessed by IHC. We determined variables associated with loco-regional recurrence (LRR), second primaries (SP), systemic recurrence (SR) and post-recurrence survival (PRS). Results: Out of 1,702 patients, 240 (14%) had an event defined as recurrence. Those with recurrent disease were significantly younger than those without,and were initially diagnosed at more advanced stages, with larger tumors, greater lymph nodal involvement and higher HG. With a median follow up of 61 months (1-250), 4.6% of patients without recurrence and 56.6% of patients with an event defined as recurrence had died. The median PRS for the LRR group was 77 months; 75 months for those who developed a SP and 22 months for patients with an SR (p <0.0001). In SR cases, the median PRS was shorter for ER- tumors than for ER+ tumors (15 vs. 26 months, respectively; p = 0.0019, HR 0.44; CI: 0.25-0.44). Conclusions: Subtype, defined through classic histopathologic parameters determined for primary tumors, was found to eb related to type of recurrence and also to prognosis after relapse.