Expression of E-cadherin and syndecan-1 in axillary lymph node metastases of breast cancer with and without extracapsular extension.

BACKGROUND: Extracapsular extension of axillary lymph node (ECE) has significantly increased the risk of locoregional and distant recurrence in breast cancer patients. OBJECTIVE: Identify markers with high biological aggressiveness since it may serve as a prognostic indicator or adjunct to standard treatment. MATERIAL AND METHOD: The authors immunostained 115 axillary lymph nodes of invasive ductal carcinoma with syndecan-1 and E-cadherin. RESULTS: The presented data shows a significantly higher number of positive lymph node (8.48 vs. 4.15; p < 0.0001) and larger primary tumor size (3.53 vs. 2.79; p = 0.0029) in ECE patients. Sixty-one cases had node positive and without evidence of ECE, 54 cases had ECE. Syndecan-1 was found to be of significantly high expression (p = 0.001). There was no significant difference in the expression of E-cadherin during progression into extracapsular area (p = 0.12). CONCLUSION: E-cadherin displays high expression in nodal breast cancer metastases that may have re-expression and has coordinate function with syndecan-1 while invading to the surrounding fatty tissue. The protein is, therefore, likely to play a role in the invasiveness and aggressiveness.

Tissue engineering of cartilage with porous polycarprolactone--alginate scaffold: the first report of tissue engineering in Thailand.

OBJECTIVE: To engineer human cartilage with porous polycaprolactone (PCL)-Alginate Scaffold. BACKGROUND: Polycaprolactone (PCL) is a prolonged degradable polymer that has good mechanical strength. The authors fabricated PCL as an ear shaped scaffold. Alginate hydrogel was used to seed chondrocyte into the PCL porous scaffold by a gel-cell seeding technique. MATERIAL AND METHOD: PCL Scaffolds were fabricated like human pinna by particle leaching technique. Chondrocyte was isolated from human rib cartilage and then cultured. The cultured chondrocyte were mixed with 1.2% alginate and b-FGF (basic-fibroblast growth factor) 5 ng/ml at a concentration of 25 x 10(6) cell/ml, then were seeded in porous PCL scaffold to make the constructs. The constructs were cultured in vitro for 1 week. Then they were implanted in subcutaneous plane of the back of six-female nude mice (5 weeks old). Two nude mice were sacrificed at 2, 3, and 6 months. Histological study was done (H&E, Alcian blue, collagen type II). RESULT: Neocartilage was formed in the porous cavity of PCL scaffold. At 2 and 3 months, neocartilage were similar to very young cartilage. At 6 months, they were mature. The delayed maturation until 6 months and the highly vascularization of neocartilage in the early phase was the effect of human b-FGF The growths of neocartilage islands in porous cavity were also observed along with degradation ofPCL inter-porous septum. CONCLUSION: This paper reports the first success of cartilage tissue engineering in Thailand.

Overexpression of c-Myc in primary central nervous system lymphoma of Thais.

BACKGROUND: c-Myc protooncogenes have been implicated in the tumourigenesis of extracerebral lymphomas, however only afew studies on this oncogenic molecule have been available for primary central nervous system lymphoma (PCNSL). OBJECTIVE: To determine the prevalence ofprotein overexpression and gene amplification of c-Myc in PCNSL and to correlate with histological and immunophenotypic subtypes of malignant lymphoma according to WHO classification of tumors of haematopoietic and lymphoid tissue 2001. SETTING: King Chulalongkorn Memorial Hospital, Thailand. DESIGN: Descriptive study. MATERIAL: 25 Thai patients presented between 2001 and 2005. METHOD: The overexpression and amplification of c-Myc in malignant lymphoma were studied by means of immunohistochemistry and chromogenic in situ hybridization (CISH), respectively, in formalin-fixed, paraffin-embedded specimens. The histomorphology and immunohistochemistry were used to subclassify PCNSLs according to WHO classification 2001. RESULTS: Fourteen males and eleven females were recruited. They were between the ages of 21 and 86 years with the mean of 53 years. Eight had documented human immune deficiency virus (HIV) infection. Four of 17 immunocompetent cases overexpressed c-Myc protein without c-Myc gene amplification. No immunocompromised cases showed overexpression of c-Myc protein. All PCNSLs were classified as diffuse large B-cell lymphoma. CONCLUSION: In PCNSL, c-Myc overexpression is notfound immunocompromised (HIV-infected) patients and is found in 23.5% of the immunocompetent individuals without c-Myc gene amplification. All PCNSLs are diffuse large B-cell lymphoma according to WHO classification 2001.

RET proto-oncogene expression of papillary thyroid carcinomas in Thai patients.

BACKGROUND: Ret proto-oncogene activation has been found in papillary thyroid carcinoma with different frequencies according to geographic location. The rate of expression ranges from 0-100 percent in the literature. This gene expression has also been studied in many Asian countries but it has never been studied in Thailand. OBJECTIVE: To study the frequency of the RET expression and their roles in predicting prognosis of papillary thyroid carcinoma among Thai patients treated at King Chulalongkorn Memorial Hospital, Bangkok, Thailand. MATERIAL AND METHOD: One hundred and one cases of papillary carcinomas were studied with immunohistochemistry for RET antibodies. All slides with routine staining were reviewed to classify cell variants and record other prognostic parameters such as size, multicentricity, extrathyroid invasion. The clinical data such as age and sex were also included for analyses. RESULTS: Forty-seven of the total 101 cases (46.5%) showed positive RET protein staining. The mean age among patients with RET negative neoplasms was 43.9 years compared with 39.8 years in RET positive group (p = 0.16). The average size of the tumors without RET expression was 2.5 cm, slightly larger than the RET positive tumors (2.1 cm)(p = 0.26). Extrathyroid invasion of the RET-positive tumors was found to be 33.2 percent while the RET negative neoplasms had 38.8 percent of this feature (p = 1). According to AMES score, the RET positive cases had only 11 percent of high-risk tumors, whereas the RET negative group comprised 23.1 percent of high-risk malignancies (p = 0.20). There was no significant difference in RET expression among cell variants (p = 1). CONCLUSION: The study of 101 papillary thyroid carcinomas at the King Chulalongkorn Memorial Hospital disclosed high frequency of RET expression (46.5%) and this is the only data among Thai patients that has ever been documented in the literature. Although, the gene expression in the tumor tends to be associated with good prognostic features but it was not distinct enough to be statistically significant.

Expression of CyclinD1, p27kp-1, and bcl-2 in plexiform neurofibroma with and without malignant transformation in neurofibromatosis type 1.

Plexiform neurofibroma (PNF) has a low potential to undergo malignant transformation. Identification of markers associated with tumor progression is important since it may serve as prognostic indicators or adjuncts to standard pathological examination. In the present study, the authors immunostained 20 neurofibromatosis type I-associated PNFs with cyclinD1, p27kip-1, and bcl-2. Six of the cases had progressed into malignant peripheral nerve sheath tumor (MPNST), and the transitional area of each sample was also stained separately in order to identify protein(s) associated with tumor progression. Cyclin D1 was found to be significantly increased in the transitional zone, compared to the ordinary PNF (p = 0.007). The protein is, thus, likely to play a role in the malignant transformation. There was no significant difference in the expression of p27kip-1 and bcl-2 during the malignant progression of PNF.

A two-phase study model for the standardization of HER2 immunohistochemical assay on invasive ductal carcinoma of the breast.

OBJECTIVES: To develop and verify a standardized protocol for HER2 immunohistochemical assays on invasive ductal carcinoma of the breast in Thailand. MATERIAL AND METHOD: A two-phase study approach was employed. In the Phase One, after verifying the proposed protocol that adopted the HercepTest procedure using readily available primary antibodies, CB11 and A0485, Lab 1 performed the HER2 immunohistochemical staining for 137 cases of invasive ductal carcinoma twice with two types of the antibody. Nine pathologists from 8 centers independently examined and scored all the 2 x 137 stained slides that were blinded for antibody type. Interobserver reliability was calculated using pair-wise kappa. Following discussion of the results, the Phase Two study was planned. Lab 2 and Lab 3 independently performed the HER2 staining according to the protocol for 60 invasive breast carcinoma cases. The same group of pathologists scored 2 x 60 stained slides that were masked for laboratories. Interobserver reliability and interlaboratory agreement from each pathologist were calculated using kappa statistics. Three interpreted categories--namely negative, equivocal and positive tests were used in the analyses. RESULTS: Phase One study showed interobserver agreement between pairs varied from kappa 0.75 (95%CI, 0.68-0.82) to 0.06 (95%CI, 0-0.14) while Phase Two study obtained pair-wise kappa scores ranged from 0.84 (95%CI, 0. 80-0.89) to 0. 65 (95%CI, 0.59-0.71). Interlaboratory kappa for each pathologist was 0.67 (95%CI, 0.61-0.73). CONCLUSION: The standardization of HER2 immunohistochemical assay was achieved through this two-phase study model. It had added benefits of improving pathologists' expertise and verifying the HER2 testing protocol to be used in Thailand.

The association of co-expression of CD44v4/MMP-9 with different nodal status in high-grade breast carcinoma patients.

BACKGROUND: Breast carcinoma is one of the most common tumors in female patients, and its metastasis is a major cause of death. An experimental model has recently found the association of CD44 with MMP-9 that facilitates tumor cell invasion and metastasis. MATERIAL AND METHOD: The CD44v4 and MMP-9 were performed on tissue in paraffin blocks of 50 cases of high-grade breast carcinoma with node positive and 50 cases with node negative. RESULTS: Increased expression of MMP-9(60%) significantly observed in high-grade breast carcinoma patients with node positive (p = 0. 004), whereas CD44v4 displays no significant difference between the two groups (p-value = 0.81). Significant co-expression of CD44v4+ / MMP-9+ (46%) was observed and correlated with node-positive patients whereas the CD44v4+ / MMP-9- (54%) express in node-negative patient (p-value = 0.01). CONCLUSION: The solely expression of CD44v4 does not associate with node status. MMP-9 plays an important role to enhance breast carcinoma cell invasion and associates with lymph node metastasis. The combined expression of CD44v4 (overexpression) and derangement of MMP-9 expression was significantly associated with nodal status.

Extranodal malignant lymphoma of the upper aerodigestive tract: prevalence of Epstien-Barr virus (EBV) infection in King Chulalongkorn Memorial Hospital.

BACKGROUND: High frequency of Epstein-Barr virus (EBV) in the normal mucosa of the upper aerodigestive tract suggests that it may serve as a reservoir for the virus. Malignant lymphomas arising in this site may be associated with EBV. OBJECTIVES: To determine the prevalence of EBV infection in extranodal malignant lymphomas of the upper aerodigestive tract. SETTING: King Chulalongkorn Memorial Hospital, Thailand. DESIGN: Descriptive study. PATIENTS: 42 Thai patients who presented between 1998 and 2003. MATERIAL AND METHOD: The expression of EBV mRNAs (EBERs) of malignant lymphoma was studied by means of in situ hybridization in formalin-fixed, paraffin-embedded specimens. RESULTS: The recruited subjects were 26 males and 16 females, and their age ranged from 3 to 85 years with the mean of 51.43 years, in 4 of them human immune deficiency virus (HIV) infection was documented. Ten of 42 cases (23.81%) expressed EBER transcripts and were extranodal NK/T-cell lymphomas, nasal type (7 cases), plasmablastic lymphomas (2 cases) and diffuse large B-cell lymphoma (1 case). Three of 4 cases (75%) of known HIV-seropositive cases were EBV-positive (2 plasmablastic lymphomas and 1 diffuse large B-cell lymphoma). CONCLUSION: In the upper aerodigestive tract, EBV was present in some but not all malignant lymphoma. It was associated with extranodal NK/T-cell lymphoma, nasal type and B-cell lymphoma arising in HIV-infected patients, but it was not found in B-cell lymphoma arising in immunocompetent patients.

Situation of laboratory service and instruments in Thailand: a descriptive study from questionnaires.

Laboratory instruments are one of the main items in laboratory investment. To establish data for the situation of laboratory service and instruments in Thailand, questionnaires were randomly sent to one hundred and twenty laboratories. Sixty-three filled questionnaires from eleven university and affiliated hospitals, thirty-four government hospitals, and eighteen private hospital laboratories were sent back to the authors to be analyzed. Only the number of samples and instruments used during office hours were analyzed in this study by descriptive method. From the data it was found that the average number of personnel and tests of the university and affiliated hospital laboratories was the highest. To analyze the efficiency of the instruments used in the laboratories, the authors compared the average service number of samples or tests to the average number of samples or tests that was calculated from the instruments. The ratio of the average number of samples or tests that were calculated from the instruments and the average service number of samples or tests for chemistry and CBC were 2.13, 3.41, 5.24 and 2.33, 2.76, 3.71 in university and affiliated hospital laboratories, government hospital laboratories, and the private hospital laboratories, respectively. From the data, it was concluded that the instrument situation in laboratories of the university and affiliated hospitals was more appropriate than government hospital and private hospital laboratories. To improve the efficiency of using laboratory instruments, more concern must be given to the management of laboratory instruments and cooperation between hospitals could increase the efficiency of the instrument investment.