Supinda Koonmee

Urinary microRNA-192 and microRNA-21 as potential indicators for liver fluke-associated cholangiocarcinoma risk group.

Opisthorchis viverrini infection induces chronic inflammation in the bile ducts, leading to periductal fibrosis (PDF), which possibly associates to cholangiocarcinoma (CCA). Patients with CCA have a poor prognosis, which is linked to asymptomatic disease and late diagnosis. Hence, detecting early stage CCA is essential. Secretory miRNAs have been promoted as biomarkers for pathological changes associated with parasitic infections, fibrosis and/or cancer. We aimed to determine levels of miR-192 and miR-21 in the urine of O. viverrini infected, periductal fibrosis (PDF) and CCA groups using qRT-PCR. We found that miR-192 was significantly higher in O. viverrini infected, PDF and also CCA groups (p<0.05) than in healthy controls. By utilizing the Receiver Operation Characteristics (ROC) analysis, miR-192 differentiated patients with opisthorchiasis (the area under the curve; AUC=0.766), PDF subjects (AUC=0.781) and CCA patients (AUC=0.682) from healthy controls. MiR-21 was significantly higher in PDF and CCA groups (p<0.05) than in healthy controls. MiR-21 discriminated PDF subjects (AUC=0.735) and CCA patients (AUC=0.682) from healthy controls. Combined levels of these two miRNAs revealed an increased AUC of 0.812 for separating opisthorchiasis, AUC of 0.815 in discriminating PDF subjects, and AUC of 0.849 in differentiating CCA from healthy controls. Odds ratios (OR) indicated high levels of miR-192/miR-21 as risk predictors for opisthorchiasis, PDF and CCA. Levels of these miRNAs declined significantly for patients following praziquantel treatment. In conclusion, urinary miR-192/miR-21 have potential as risk indicators for opisthorchiasis and PDF-associated CCA in the endemic region.

Molecular identification of a causative parasite species using formalin-fixed paraffin embedded (FFPE) tissues of a complicated human pulmonary sparganosis case without decisive clinical diagnosis

PCR-based molecular diagnosis was made for the identification of causative agents of the clinically suspected pulmonary proliferative sparganosis case found in Thailand using formalin-fixed paraffin-embedded (FFPE) biopsy specimens. As a reference, FFPE biopsy specimen from a typical cutaneous sparganosis case was examined together. DNA samples were extracted from tissues and two partial fragments of cytochrome c oxidase subunit 1 (cox1) gene were amplified for the detection of Spirometra DNA. Two cox1 fragments were amplified successfully for both specimens. After alignment of nucleotide sequences of the PCR-amplicons, the causative agents of both cases were identified as Spirometra erinaceieuropaei.

Anal Cancer Screening by Modified Liquid-Based Cytology in an HIV Clinic

This study aimed to screen for anal cancer and to determine its cytomorphology using liquid-based cytology (LBC) with specimens preserved in 95% ethyl alcohol. Anal swabs were collected for cytological examination from 177 adult, HIV-infected patients. After collection, sample slides were reviewed and classified according to their cytomorphology using the modified Bethesda 2001 system. An abnormal anal Pap smear was found in 26.0% of the patients. The diagnoses were: 66.7% negative for intraepithelial lesions (NIL), 14.1% with atypical squamous cells of undetermined significance (ASC-US), 10.7% (19) with low-grade squamous intraepithelial lesions (LSIL), and 1.13% with high-grade squamous intraepithelial lesions (HSIL). The cytological evaluation was an unsatisfactory result only with 6.67%. The present modified LBC using 95% ethyl alcohol as the preservative could thus be used for anal cancer screening. The number of SILs in Thai HIV-infected patients is lower than that in Western countries. We found anal cytology a satisfactory tool for early screening and detection of anal dysplasia commonly found in high-risk, HIV-infected patients.

Extramedullary hematopoiesis in pyogenic granuloma

Extramedullary hematopoiesis (EMH) suggests the presence of hematopoietic stem cells (HSC) outside bone marrow. EMH has been reported, albeit rarely, in pyogenic granuloma (PG), a polypoid lobular capillary hemangioma. However, statistical data have hitherto been lacking on the actual incidence of EMH in PG. Therefore, we here reviewed 157 consecutive cases using routine diagnostic surgical slides and found unequivocal EMH in 17 (10.8%). This indicates that EMH is a rather common finding in PG, which could thus have strong potential to be an important resource for the study of HSC.

Upregulation of endothelial nitric oxide synthase (eNOS) and its upstream regulators in Opisthorchis viverrini associated cholangiocarcinoma and its clinical significance

Endothelial nitric oxide synthase (eNOS) is an isoform of the enzyme nitric oxide synthase (NOS) which is constitutively expressed in endothelial cells and plays important roles in vasodilation. We previously reported the importance of eNOS activation in cholangiocarcinoma (CCA) tissues and cell lines. The present study aims to investigate the relative abundance of eNOS and phosphorylated eNOS (P-eNOS) and their upstream regulators VEGFR3, VEGFC, EphA3 and ephrin-A1, in the Opisthorchis viverrini (Ov)/N-nitrosodimethylamine (NDMA)-induced hamster CCA model and in human CCA by semiquantitative immunohistochemical analysis of the relevant tissues. Results from the hamster model suggested an increase in eNOS and P-eNOS and upstream regulators during CCA genesis. In human CCA, high immunohistochemical staining intensity of all investigated proteins was associated with the presence of metastasis. A pairwise analysis of the staining data for eNOS and its upstream regulators showed that a concurrent increase in eNOS/VEGFR3, eNOS/ephrin-A1, eNOS/VEGFC and eNOS/EphA3 was significantly associated with metastasis. An increase in eNOS/VEGFR3, eNOS/ephrin-A1 was also associated with non-papillary type CCA. Additionally, an increase in eNOS and P-eNOS was significantly correlated with a high micro-vessel level (P=0.04). Our results indicate that the development of CCA involves upregulation of eNOS and P-eNOS and their regulators. This may drive angiogenesis and metastasis in CCA.

Joinpoint analysis trends in the incidence of colorectal cancer in Khon Kaen, Thailand (1989 - 2012)

Objective: This study focused on recent changes in the incidence of colorectal cancer (CRC) in Khon Kaen, Thailand. Methods: Data for CRC over the period 1989 to 2012 from the population-based cancer registry of Khon Kaen province were employed. Age-standardized incidence rates (ASR) were calculated and classified into 4 age-groups for comparison. Joinpoint regression analysis was used to detect changes in trends among each line segment and an overall line was generated, whether increasing or decreasing, with annual percent change (APC) and average annual percent change (AAPC). Results: There were 3,364 CRC cases included in the analysis, 72.2% histological confirmed and 53.5% in men. Trends of ASRs generally demonstrated gradual increase over the period 1989 to 2012. For those aged under 45 or 50 years there was slight overall increase, with a somewhat zigzag pattern. From joinpoint analysis, the trends of all aged groups were found to be increasing among both men and women: aged 45 years and older group AAPC=3.40, 2.30 and 3.90, respectively); aged 50 years and older group AAPC=2.90, 2.20 and 3.40; aged under 45 years AAPC=6.30, 6.00 and 6.90; and aged under 50 years (AAPC=5.70, 3.20 and 5.70. Conclusions: ASRs for CRC have been gradually increasing in the northeast region of Thailand. Future studies should consider the subsite distribution.

Breast Cancer Incidence Trends and Projections in Northeastern Thailand

Background The northeast has the lowest incidence of breast cancer of all regions in Thailand, although national rates are increasing. The heterogeneity in subnational trends necessitates a comprehensive evaluation of breast cancer incidence trends and projections to provide evidence for future region-specific strategies that may be employed to attenuate this growing burden. Methods Joinpoint regression and age-period-cohort modeling were used to describe trends from 1988–2012. Data was projected from three separate models to provide a range of estimates of incidence to the year 2030 by age group. Results Age-standardized rates (ASRs) increased significantly for all women from 1995–2012 by 4.5% per year. Rates for women below age 50 increased by 5.1% per year, while women age 50 years and older increased by 6% per year from 1988–2012. Projected rates show that women age 50 years and older have the largest projected increase in ASRs by 2030 compared to younger women and all women combined. Conclusions Breast cancer trends in Khon Kaen are presently lower than other regions but are expected to increase and become comparable to other regions by 2030, particularly for women ages 50 years and older.

Evaluating a preoperative protocol that includes magnetic resonance imaging for lymph node metastasis in the Cholangiocarcinoma Screening and Care Program (CASCAP) in Thailand

BackgroundTreatment planning especially liver resection in cholangiocarcinoma (CCA) depends on the extension of tumor and lymph node metastasis which is included as a key criterion for operability. Magnetic resonance imaging (MRI) offers a rapid and powerful tool for the detection of lymph node metastasis (LNM) and in the current manuscript is assessed as a critical tool in the preoperative protocol for liver resection for treatment of CCA. However, the accuracy of MRI to detect LNM from CCA had yet to be comprehensively evaluated.MethodsThe accuracy of MRI to detect LNM was assessed in a cohort of individuals with CCA from the Cholangiocarcinoma Screening and Care Program (CASCAP), a screening program designed to reduce CCA in Northeastern Thailand by community-based ultrasound (US) for CCA. CCA-positive individuals are referred to one of the nine tertiary centers in the study to undergo a preoperative protocol that included enhanced imaging by MRI. Additionally, these individuals also underwent lymph node biopsies for histological confirmation of LNM (the “gold standard”) to determine the accuracy of the MRI results.ResultsMRI accurately detected the presence or absence of LNM in only 29 out of the 51 CCA cases (56.9%, 95% CI 42.2–70.7), resulting in a sensitivity of 57.1% (95% CI 34.0–78.2) and specificity of 56.7% (95% CI 37.4–74.5), with positive and negative predictive values of 48.0% (95% CI 27.8–68.7) and 65.4% (95% CI 44.3–82.8), respectively. The positive likelihood ratio was 1.32 (95% CI 0.76–2.29), and the negative likelihood ratio was 0.76 (95% CI 0.42–1.36).ConclusionsMRI showed limited sensitivity and a poor positive predictive value for the diagnosis of LNM for CCA, which is of particular concern in this resource-limited setting, where simpler detection methods could be utilized that are more cost-effective in this region of Thailand. Therefore, the inclusion of MRI, a costly imaging method, should be reconsidered as part of protocol for treatment planning of CCA, given the number of false positives, especially as it is critical in determining the operability for CCA subjects.

Predictive relevance of tumor-infiltrating lymphocytes in breast cancer

&NA; Tumor-infiltrating lymphocytes (TILs) are considered to be a manifestation of the host anti-tumor response. We aim to evaluate the prognostic significance of the immunophenotype, density and distribution of TILs in breast cancer samples. This was a retrospective study, using paraffin-embedded samples obtained from 43 breast cancer patients, treated at Mahasarakham Hospital between 2012 and 2013. TILs and prognostic markers were evaluated by immunohistochemical staining of tissue microarray cores, employing monoclonal antibodies to lymphocyte markers (i.e., CD3, CD4, CD8 and CD20) and markers for breast cancer (i.e., ER, PR, HER2 and Ki-67). TILs were categorized into: 1) intratumoral lymphocytes (ITLs) when found within tumor cell nests), and 2) peritumoral lymphocytes (PTLs) when infiltrating into the stroma, adjacent to the invasive front of tumor. Results: A positive correlation was found between high density CD8+ PTLs and breast carcinoma without lymph node metastasis (p = 0.044), and an inverse correlation was found between the density of CD4+ PTLs and estrogen receptor expression (p = 0.027). The density of CD8+ PTLs as well as the number of CD20+ ITLs were independently, positively correlated with tumor size.