Hsiao-Lin Hwa

Mutation spectrum of the connexin 26 (GJB2) gene in Taiwanese patients with prelingual deafness

Purpose: To determine the mutation spectrum of the connexin 26 gene among 324 Taiwanese patients with prelingual deafness and the carrier rate of gene mutation in another 432 unrelated control subjects.Methods: The coding region of the connexin 26 gene was sequenced in both directions to detect mutation in all 756 samples.Results: Among the 756 samples tested, 21 connexin 26 variants were detected, including 7 novel ones. The 235delC mutation was the most common, accounting for 57.6% of the mutant alleles. Among patients, 48 (14.8%) had connexin 26 gene mutations. In the control group, the carrier rate of connexin 26 mutation was estimated at 2.8%.Conclusion: The mutation spectrum of the connexin 26 gene is wide, with more than half of the patients having only one mutation detected. Thus, further efforts are needed to look for possible existence of a second mutant allele.

Expression of the messenger RNA for gonadotropin-releasing hormone and its receptor in human cancer cell lines

The presence of gonadotropin-releasing hormone (GnRH) binding sites in biopsy samples of human epithelial ovarian cancer and ovarian tumor cell lines as well as the demonstration of the inhibitory effects of GnRH analogues on the growth of these cells raised the possibility that GnRH is produced locally by ovarian cancer cells. In order to investigate an autocrine/paracrine regulatory mechanism in human carcinomas, we have studied the expression of GnRH and GnRHR mRNA in human ovarian epithelial cell lines (OVCAR-3 and SKOV-3), human choriocarcinoma cell line (JEG-3) and human hepatocarcinoma cell line (HepG 2). Using primers corresponding to published human GnRH and GnRHR cDNA sequences, predicted PCR products were obtained from these cell lines by reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by Southern hybridization. Sequencing analysis of GnRH PCR products showed that their sequences have 100% identity to the published human GnRH cDNA sequence. These results indicated that GnRH and GnRHR genes are expressed in all the cell lines tested in the present study, and strengthen the concept that GnRH may act as an autocrine regulator on the growth of cancer cells.

Effect of Pre-Eclampsia–Eclampsia on Major Cardiovascular Events Among Peripartum Women in Taiwan

There is no large-scale population-based study to clarify the association between major adverse cardiovascular events (MACEs) and pre-eclampsia/eclampsia. A population-based Taiwanese cohort study was performed in 1,132,064 parturients from 1999 to 2003 using a dataset linking birth certificates and National Health Insurance hospital discharge data. Sociodemographic factors and obstetric complications were used in multivariate logistic regression models to determine adjusted hazard ratios of pre-eclampsia/eclampsia on risks of MACEs and mortality during pregnancy to at least the third year postpartum. Incidence rates of MACEs and all maternal mortality in women with pre-eclampsia/eclampsia were 16.21 and 40.38 per 100,000 patients per year, respectively. Women with pre-eclampsia/eclampsia had a 13.0-fold higher incidence of myocardial infarction, a 8.3-fold higher incidence of heart failure, a 14.5-fold higher incidence of stroke, a 12.6-fold higher incidence of MACEs, a 7.3-fold higher incidence of MACEs without stroke, a 2.3-fold higher incidence of MACE-related deaths, and a 6.4-fold higher incidence of overall death than women without pre-eclampsia/eclampsia. Kaplan-Meier survival curve discriminated in MACEs, nonstroke MACEs, MACE related death and overall death. In conclusion, women with pre-eclampsia/eclampsia have a significantly higher risk of MACEs, especially myocardial infarction and stroke, during pregnancy and their risk remains significant to ≥36 months postpartum. Our results suggest that women with pre-eclampsia/eclampsia should be closely monitored during pregnancy and for up to ≥3 years postpartum.

Molecular genetic study of Pompe disease in Chinese patients in Taiwan

Pompe disease is caused by mutations in the acid α‐glucosidase (GAA) gene. Multiple kinds of mutations in the GAA gene have been reported worldwide. In order to elucidate the molecular basis of the disease in Taiwanese patients of Chinese origin, we have recruited 11 unrelated families who had at least one member with Pompe disease for study. We used 16 pairs of oligonucleotide primers to amplify all the coding regions from exon 2 to 20 in the family members. The coding regions were sequenced on both the sense and antisense strands. We identified 7 different mutations in 17 alleles but failed to identify the defects in the other 5 alleles. The most common defect was D645E (Asp645Glu), accounting for 36% (8/22 alleles) of mutations, followed by G615R (Gly615Arg) (3 alleles); 1411del4 (Glu471‐shift) (2 alleles); and one allele each of R600H (Arg600His); ΔN675 (ΔAsn675); 2380delC (Arg794‐shift) and 2815delGT (Val939‐shift). The molecular defects of Pompe disease are highly heterogeneous in Chinese. Characterization of the molecular defects of the disease is useful for a genotype‐phenotype correlation and for genetic counseling and prenatal diagnosis. Hum Mutat 13:380–384, 1999.

The Interactions Between GPR30 and the Major Biomarkers in Infiltrating Ductal Carcinoma of the Breast in an Asian Population

OBJECTIVE G-protein-coupled receptor 30 (GPR30) has been reported to be a novel estrogen receptor alpha (ERalpha) in vitro. Therefore, the interactions among GPR30, ERalpha, progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER-2/neu), and their prognostic utilities in the infiltrating ductal carcinoma (IDC) of the breast were evaluated. MATERIALS AND METHODS Messenger RNA (mRNA) levels of GPR30, ERalpha, PR and HER-2/neu in the tumor samples of 118 Taiwanese IDC patients and 27 non-tumor mammary tissues were measured via quantitative polymerase chain reaction analyses. The correlations of GPR30 mRNA levels with clinical parameters, i.e. tumor/non-tumor, ERalpha, PR, HER-2/neu, age, lymph node metastasis, lymph-vascular invasion, grade, stage and patient survival, were assessed by using appropriate statistical analyses. RESULTS GPR30 expression was observed to be lower in IDC (p < 0.001) than in non-tumor mammary tissues. Importantly, GPR30 mRNA level was positively correlated with that of ERalpha (p = 0.001) and PR (p = 0.001) but not correlated with that of HER-2/neu when they were analyzed as continuous variables. However, lower GPR30 was noticed in tumors with HER-2/neu protein overexpression. GPR30 expression was not correlated with age, lymph node metastasis, lymph-vascular invasion, grade and stage in IDC. GPR30 expression was not an independent prognostic factor for patient survival. CONCLUSION GPR30 expression is downregulated in IDC. GPR30 is preferentially co-expressed with ER and/or PR but is lowly expressed in HER-2/neu(+) tumors. The correlation of GPR30 expression with clinical parameters, including patient survival, was not evident in this cohort.

Correlation of Prenatal Ultrasound and Postnatal Outcome in Meconium Peritonitis

Objectives: To study the relationship between prenatal ultrasound features and postnatal course of meconium peritonitis. Methods: Meconium peritonitis was diagnosed by prenatal ultrasound. Fetuses were treated by intrauterine paracentesis of ascites when indicated, and symptomatic newborns received surgery. Results: Totally 17 cases were enrolled. Prenatal ultrasound findings include abdominal calcification (16/17), fetal ascites (12/17), hydramnios (9/17), pseudocyst (7/17) and dilated bowel loop (6/17). Persistent ascites, pseudocyst or dilated bowel loop are most sensitive (92%) to predict postnatal surgery (p = 0.022). The survivors have a higher gestational age at birth (36.4 vs. 33.3 weeks, p = 0.008). Persistent ascites and postnatal persistent pulmonary hypertension of the newborns significantly correlate with neonatal mortality (p = 0.029 and 0.022). Conclusion: Prenatal ultrasound can predict the neonatal outcome in meconium peritonitis.

SPARC (Osteonectin) in Breast Tumors of Different Histologic Types and Its Role in the Outcome of Invasive Ductal Carcinoma

Abstract:  The purpose of this study was to characterize the immunohistochemical distribution of secreted protein acidic and rich in cystein (SPARC) in benign and malignant breast tumors of different histologic types and define its association with the outcome of invasive ductal carcinoma (IDC) patients. A total of 286 samples of benign and malignant breast lesions between 1994 and 2005 were retrieved from National Taiwan University Hospital. Up to 11 years clinical follow‐up data were available for 185 patients with IDC. Immunohistochemistry staining with SPARC was performed in tissue microarray or whole section. The association of expression of SPARC and cumulative overall survival of IDC patients were analyzed using Kaplan–Meier survival analysis and Cox regression analysis. Secreted protein acidic and rich in cystein was not expressed in benign breast phylloides and all benign breast tumors, while expressed in 17.2% of IDC, 85% of metaplastic carcinoma of the breast (MCB), and all malignant breast phylloides. Secreted protein acidic and rich in cystein was strongly expressed in mesenchymal components of MCB and expression levels in epithelial components were variable. The correlation of positive expression of SPARC and poor long‐term survival in IDC is significant (p = 0.004). Individuals with positive SPARC expression had 2.34 times higher hazard of death compared with those with negative SPARC expression after adjusting for factors including positive lymph node, TNM tumor stage, estrogen receptor, and progesterone receptor. Secreted protein acidic and rich in cystein may be useful as a prognostic indicator for IDC.

Thirteen X-chromosomal short tandem repeat loci multiplex data from Taiwanese

Study results of variations in the X chromosome are useful tools in researching the genetic diversity of human populations and individual identification. We developed a 13 X chromosomal short tandem repeat (STR) multiplex system (DXS6807, DXS8378, DSX9902, DXS7132, DXS9898, DXS6809, DXS6789, DXS7424, DXS101, GATA172D05, HPRTB, DXS8377, DXS7423) amplified in one single polymerase chain reaction. DNA samples of 113 male and 108 female Taiwanese Han subjects were successfully analyzed using this 13 X-STR multiplex system. The distributions of allele frequencies were examined for independence. DXS8377, DXS101, DXS6789, and DXS6809 were found to be the most polymorphic markers in this study. High values of discrimination power and mean exclusion chance without significant evidence of association between these markers were obtained. In conclusion, this 13 X chromosomal STR multiplex system offers considerable forensic efficiency and may be useful in forensic identification casework.

TNF-related apoptosis-inducing ligand (TRAIL) induces osteoclast differentiation from monocyte/macrophage lineage precursor cells

Human osteoclast formation from mononuclear phagocyte precursors involves interactions between tumor necrosis factor (TNF) ligand superfamily members and their receptors. Many of the proinflammatory cytokines and growth factors implicated in inflammatory processes have also been demonstrated to impact osteoclast differentiation and function. Recent evidence indicates that the TNF-related apoptosis-inducing ligand (TRAIL) of the TNF ligand superfamily, which was initially thought to induce apoptosis in many transformed cell lines, can serve as an effector molecule in activated T cells. We show in this work that TRAIL can induce osteoclast formation from human monocytes and murine RAW264.7 macrophages. We demonstrated that both cell models differentiate into osteoclast-like cells in the presence of TRAIL in a dose-dependent manner, as evaluated in terms of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells and bone resorption activity. The TRAIL-induced osteoclast differentiation is independent of caspase activation and apoptosis induction activity. However, TRAIL-induced osteoclastogenesis is dependent on activation of NF-kappaB, ERK, and p38 MAP kinase. Thus, our data demonstrate that TRAIL induces osteoclast differentiation via direct engagement with the TRAIL death receptor through a signaling pathway distinct from apoptosis. Our results indicate that in addition to triggering apoptosis, TRAIL induces osteoclast differentiation. It provides a novel role for TRAIL in regulating osteoclast differentiation and in osteoimmunology.

Prediction of breast cancer and lymph node metastatic status with tumour markers using logistic regression models

AIMS Early detection of breast cancer can improve disease mortality. The aim of this study was to evaluate the effectiveness of serum biomarkers in the detection of primary breast cancer and lymph node metastatic status. METHODS Serum samples were obtained from 55 female patients with breast cancer and 39 women without breast cancer. For these subjects, clinicopathological data were collected and serum levels of carcinoembryonic antigen, breast cancer-specific cancer antigen 15.3 (CA15-3), tissue polypeptide-specific antigen (TPS), soluble interleukin-2 receptor (sIL-2R) and insulin-like growth factor binding protein-3 (IGFBP-3) were assayed. Univariate and multivariate logistic regression were performed to evaluate the association between biomarkers and breast cancer, as well as lymph node metastatic status. RESULTS For breast cancer prediction, the serum level of TPS had the best predictive value, with a sensitivity of 80% at an optimal cut-off value of 69.1 U L(-1). The combination of TPS, CA15-3 and IGFBP-3 with logistic regression model increased the sensitivity to 85%. For lymph node metastasis prediction, the serum level of sIL-2R had the best predictive value, with a sensitivity of 66% at an optimal cut-off value of 286 U mL(-1). The combination of sIL-2R and TPS with logistic regression model increased the sensitivity to 69%. CONCLUSION TPS may be useful in the detection of primary breast cancer, while sIL-2R may be useful in lymph node metastasis prediction. The combination of more than one biomarker with logistic regression model can improve the predictive sensitivity.