Chia-Hsun Hsieh

A negative selection system PowerMag for effective leukocyte depletion and enhanced detection of EpCAM positive and negative circulating tumor cells

BACKGROUND Circulating tumor cells (CTCs) can be enriched from peripheral blood by leukocyte depletion. In this study, a PowerMag system was designed to enhance leukocyte depletion and CTC detection. METHODS The efficiency of PowerMag in leukocyte depletion and recovery of CTCs was evaluated. The cell populations in leukocyte-depleted cell filtrate were characterized by immunofluorescence staining using anti-EpCAM and anti-CD45 antibodies. The suitability of PowerMag in monitoring cancer patient CTCs was assessed by analyzing blood samples collected at baseline and 2 weeks after baseline from the patients with colorectal cancer (CRC, n=24) and head and neck squamous cell carcinoma (HNSCC, n=28). RESULTS Only few contaminated CD45(+)-leukocytes were retained in the leukocyte-depleted cell filtrate. The enriched CTCs were viable with a recovery rate of 46-62%. Immunofluorescence staining of the nucleated cells enriched by PowerMag revealed 2 major cell populations EpCAM(+)CD45(-) and EpCAM(-)CD45(-). The number for both cell types was significantly increased in cancer patients when compared to healthy control. After chemotherapy, the number of EpCAM(+)CD45(-) and EpCAM(-)CD45(-) cells was decreased more prominently for CRC and HNSCC patients, respectively. CONCLUSIONS PowerMag holds great promise as a platform for leukocyte depletion and CTC detection in a clinical setting.

NITRIC OXIDE AND INTERLEUKIN-1β MEDIATE NORADRENERGIC INDUCED CORTICOTROPHIN-RELEASING HORMONE RELEASE IN ORGANOTYPIC CULTURES OF RAT PARAVENTRICULAR NUCLEUS

Noradrenergic inputs from the brainstem are critical for the central stress response. It has been suggested that endogenous interleukin-1beta (IL-1beta) is involved in norepinephrine (NE)-induced release of corticotropin-releasing hormone (CRH) from the paraventricular nucleus of the hypothalamus (PVN). However, no IL-1 receptor on PVN CRH neurons has been identified. Therefore we hypothesized that the action of IL-1beta in the PVN requires downstream modulators that eventually lead to CRH release by PVN neurons. In the current study, we used organotypic cultures from neonatal rat PVN which display neuroendocrine characteristics suitable for in vitro studies. Pharmacological treatments with NE or IL-1beta elicited nitric oxide (NO) release from the PVN cultures, implying that local NO might be a candidate for modulating the action of IL-1beta. In addition, NE treatments significantly increased IL-1beta and CRH release. Treatment with IL-1beta or sodium nitroprusside also induced CRH release. Next, we also showed that either an IL-1 receptor antagonist or NOS inhibitor Nomega-nitro-L-arginine (L-NNA) attenuated the NE-induced CRH release. These results suggest that IL-1beta and NO are involved in NE-induced CRH release. Moreover, we found that application of L-NNA attenuated IL-1beta-induced CRH release, indicating that NO likely mediates this process. In summary, the current study demonstrates that IL-1beta plays a significant role in NE-induced CRH release, and that neuroendocrine response in the PVN may depend on local NO action.

Validation of a Palliative Prognostic Index to Predict Life Expectancy for Terminally Ill Cancer Patients in a Hospice Consultation Setting in Taiwan

BACKGROUND The aim of our study was to assess the practical utility of the palliative prognostic index (PPI) as a prognostic tool used by nurse specialists in a hospice consultation setting in Taiwan. METHODS In total, 623 terminal cancer patients under hospice consultation care from one medical center in northern Taiwan were enrolled between January 1 and June 30, 2011. PPI was assessed by a nurse specialist at first hospice consultation and patients categorized into groups by prognosis (good, intermediate, poor). Patient survival was analyzed retrospectively to determine significance of between-group differences. RESULTS By PPI sum score, 37.2% of patients were in the good prognosis group, 18% in the intermediate prognosis group and 44.8% in the poor prognosis group. The death rates were 56%, 81.2% and 89.6% and median survivals were 76, 18 and 7 days, respectively. The hazard ratio was 0.19 (95% confidence interval [CI] 0.10-0.24, p<0.001) for the poor versus good prognosis group and 0.54 (95% CI 0.43-0.69, p<0.001) for the poor versus intermediate prognosis group. The sensitivity and specificity for the poor prognosis group was 66% and 71%; the positive predictive value and negative predictive value were 81% and 52%, respectively, to predict patient death within 21 days (area under the curve of the receiver operating characteristic was 0.68). CONCLUSIONS Assessment by PPI can accurately predict survival of terminal cancer patients receiving hospice consultation care. PPI is a simple tool and can be administered by nurse members of hospice consultation teams.

Clinical scenario of EBV DNA follow-up in patients of treated localized nasopharyngeal carcinoma

OBJECTIVE In this study, we investigated the usefulness and limitations of EBV-DNA follow-up in patients who had treated localized nasopharyngeal carcinoma. METHODS Study subjects comprised 389 patients who had received treatment for localized nasopharyngeal carcinoma in our department. Copy numbers of EBV-DNA in plasma were assessed by real-time quantitative PCR. Patients in whom disease recurrence was suspected underwent image evaluation, esp. PET scan, and tissue proof if it is feasible. Lesions of undermined nature were confirmed by sequential follow-up. RESULTS Plasma EBV-DNA was detectable in 60 of 63 (95%) patients with metastatic disease and all had positive PET findings. In addition, of the 45 patients with localized recurrent disease, plasma EBV-DNA was detectable in 23 (51%) patients and positive PET scan results were obtained in 40 (89%) of the patients. Of the 284 patients who were disease free, plasma EBV-DNA was detected in 90 (32%) patients. Of the 19 patients in disease free group who were suspected disease recurrence receiving PET scanning, 7 positive PET images were found including 3 second primary malignancy and 4 non-cancer lesions. Two lymphoma cases with positive EBV-DNA value sequentially attacked before or after their NPC were diagnosed. With the cutoff value of 400copies/ml of EBV-DNA, the positive predict value was 73.5% and the negative predict value was 82.1%. The sensitivity was 0.46 and the specificity was 0.94. CONCLUSIONS EBV-DNA was a good marker for detecting metastatic failure in treated localized NPC. However, careful interpretation with complements from image examination was needed for locoregional failure and other false positive or false negative situations.

Characteristics of patients with hematologic malignancies who received palliative care consultation services in a medical center.

This study aimed to compare the characteristics of patients with hematologic malignancies and solid cancers who received palliative care. A total of 124 patients with hematologic malignancy and 3032 patients with solid cancer, who received palliative care consultation services between 2006 and 2010 in a medical center in Taiwan, were retrospectively analyzed. Higher prevalence of oral stomatitis, diarrhea, and hematologic symptoms including infection, fever, severe anemia, and bleeding, and lower prevalence of constipation, abdominal distension, and pain were observed in patients with hematologic malignancies compared to that in patients with solid cancer. The interval from hospital admission to palliative care referral was longer for patients with hematologic malignancy than that for patients with solid cancer. Hematologists should refer patients earlier, and palliative care specialists should understand the specific needs of patients with hematologic malignancy.

Development of a Microfluidic-Based Optical Sensing Device for Label-Free Detection of Circulating Tumor Cells (CTCs) Through Their Lactic Acid Metabolism

This study reports a microfluidic-based optical sensing device for label-free detection of circulating tumor cells (CTCs), a rare cell species in blood circulation. Based on the metabolic features of cancer cells, live CTCs can be quantified indirectly through their lactic acid production. Compared with the conventional schemes for CTC detection, this label-free approach could prevent the biological bias due to the heterogeneity of the surface antigens on cancer cells. In this study, a microfluidic device was proposed to generate uniform water-in-oil cell-encapsulating micro-droplets, followed by the fluorescence-based optical detection of lactic acid produced within the micro-droplets. To test its feasibility to quantify cancer cells, experiments were carried out. Results showed that the detection signals were proportional to the number of cancer cells within the micro-droplets, whereas such signals were insensitive to the existence and number of leukocytes within. To further demonstrate its feasibility for cancer cell detection, the cancer cells with known cell number in a cell suspension was detected based on the method. Results revealed that there was no significant difference between the detected number and the real number of cancer cells. As a whole, the proposed method opens up a new route to detect live CTCs in a label-free manner.

Trajectory and predictors of quality of life during the dying process: roles of perceived sense of burden to others and posttraumatic growth

PurposeQuality of life (QOL) at end of life (EOL) is related to important themes, e.g., “sense of burden to others” and “perceived posttraumatic growth,” which have never been investigated concurrently. The purposes of this study were: (1) to describe the trajectory of QOL during the dying process and (2) to identify determinants of QOL, including the roles of perceived sense of burden to others and posttraumatic growth.MethodsA convenience sample of 313 terminally ill cancer patients was surveyed and longitudinally followed until death. QOL was measured by a modified McGill quality of life scale, and determinants were evaluated by a multiple linear regression model with the generalized estimating equation.ResultsTerminally ill Taiwanese cancer patients’ QOL decreased substantially as their death approached. However, after controlling for confounders, patients’ QOL did not change significantly in the last months of their life. QOL was significantly better for female and non-middle-aged patients with a religious affiliation of Buddhism/Taoism. Poorer QOL tended to be experienced by patients with greater physical symptom distress, anxiety, and depression. Patient QOL increased with greater tangible support, but decreased with greater emotional and affectionate social support. QOL was diminished by a greater degree of perceived burden to others but improved with greater perceived posttraumatic growth.ConclusionsDeteriorating QOL as death approaches may not be inevitable. Optimal QOL at EOL may be achieved by interventions designed to adequately manage physical and psychological symptoms, enhance social support, lighten perceived sense of burden to others, and facilitate experiences of posttraumatic growth.

Application of optically-induced-dielectrophoresis in microfluidic system for purification of circulating tumour cells for gene expression analysis- Cancer cell line model.

Circulating tumour cells (CTCs) in a blood circulation system are associated with cancer metastasis. The analysis of the drug-resistance gene expression of cancer patients’ CTCs holds promise for selecting a more effective therapeutic regimen for an individual patient. However, the current CTC isolation schemes might not be able to harvest CTCs with sufficiently high purity for such applications. To address this issue, this study proposed to integrate the techniques of optically induced dielectrophoretic (ODEP) force-based cell manipulation and fluorescent microscopic imaging in a microfluidic system to further purify CTCs after the conventional CTC isolation methods. In this study, the microfluidic system was developed, and its optimal operating conditions and performance for CTC isolation were evaluated. The results revealed that the presented system was able to isolate CTCs with cell purity as high as 100%, beyond what is possible using the previously existing techniques. In the analysis of CTC gene expression, therefore, this method could exclude the interference of leukocytes in a cell sample and accordingly contribute to higher analytical sensitivity, as demonstrated in this study. Overall, this study has presented an ODEP-based microfluidic system capable of simply and effectively isolating a specific cell species from a cell mixture.

Cancers Screening in an Asymptomatic Population by Using Multiple Tumour Markers.

Background Analytic measurement of serum tumour markers is one of commonly used methods for cancer risk management in certain areas of the world (e.g. Taiwan). Recently, cancer screening based on multiple serum tumour markers has been frequently discussed. However, the risk–benefit outcomes appear to be unfavourable for patients because of the low sensitivity and specificity. In this study, cancer screening models based on multiple serum tumour markers were designed using machine learning methods, namely support vector machine (SVM), k-nearest neighbour (KNN), and logistic regression, to improve the screening performance for multiple cancers in a large asymptomatic population. Methods AFP, CEA, CA19-9, CYFRA21-1, and SCC were determined for 20 696 eligible individuals. PSA was measured in men and CA15-3 and CA125 in women. A variable selection process was applied to select robust variables from these serum tumour markers to design cancer detection models. The sensitivity, specificity, positive predictive value (PPV), negative predictive value, area under the curve, and Youden index of the models based on single tumour markers, combined test, and machine learning methods were compared. Moreover, relative risk reduction, absolute risk reduction (ARR), and absolute risk increase (ARI) were evaluated. Results To design cancer detection models using machine learning methods, CYFRA21-1 and SCC were selected for women, and all tumour markers were selected for men. SVM and KNN models significantly outperformed the single tumour markers and the combined test for men. All 3 studied machine learning methods outperformed single tumour markers and the combined test for women. For either men or women, the ARRs were between 0.003–0.008; the ARIs were between 0.119–0.306. Conclusion Machine learning methods outperformed the combined test in analysing multiple tumour markers for cancer detection. However, cancer screening based solely on the application of multiple tumour markers remains unfavourable because of the inadequate PPV, ARR, and ARI, even when machine learning methods were incorporated into the analysis.

Impact of high self‐perceived burden to others with preferences for end‐of‐life care and its determinants for terminally ill cancer patients: a prospective cohort study

Self‐perceived burden to others (SPB) is a major concern of terminally ill cancer patients and is frequently factored into end‐of‐life (EOL) care decision‐making. However, changes in and determinants of SPB and its longitudinal impact on preferences for EOL care over the dying process have not been investigated. Our study was aimed at filling this gap in knowledge.