Differentiating between dengue fever and malaria using hematological parameters in endemic areas of Thailand.

BACKGROUND: Dengue fever (DF) and malaria are the two major public health concerns in tropical countries such as Thailand. Early differentiation between dengue and malaria could help clinicians to identify patients who should be closely monitored for signs of dengue hemorrhagic fever or severe malaria. This study aims to build knowledge on diagnostic markers that are used to discriminate between the infections, which frequently occur in malaria-endemic areas, such as the ones in Thailand. METHODS: A retrospective study was conducted in Phop Phra Hospital, a hospital located in the Thailand-Burma border area, a malaria-endemic area, between 2013 and 2015. In brief, data on 336 patients infected with malaria were compared to data on 347 patients infected with DF. RESULTS: White blood cells, neutrophil, monocyte, eosinophil, neutrophil-lymphocyte ratio, and monocyte-lymphocyte ratio were significantly lower in patients with DF compared to patients with malaria (P < 0.0001). In contrast, red blood cells, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration were significantly higher in patients with DF as compared to patients with malaria (P < 0.0001). A decision tree model revealed that using neutrophils, lymphocyte, MCHC, and gender was guided to discriminate between dengue and malaria infection. CONCLUSION: This study concluded that several hematological parameters were different for diagnosing DF and malaria. A decision tree model revealed that using neutrophils, lymphocyte, MCHC, and gender was guided to discriminate patients with dengue and malaria infection. In addition, using these markers will thus lead to early detection, diagnosis, and prompt treatment of these tropical diseases.

Establishment and characterization of a novel human cholangiocarcinoma cell line with high metastatic activity.

Cholangiocarcinoma (CCA) is a highly metastatic tumor, and the lung is a common site of metastasis. A greater understanding of the biology of metastases is needed to improve treatment outcomes. Herein, a highly metastatic human CCA subline, KKU-213L5 from an original cell line, KKU-213 that has marginally metastatic ability, was established and characterized. KKU-213L5 was selected in vivo through the fifth serial passage of pulmonary metastasized tissues via tail-vein injection in NOD/scid/Jak3 mice. The metastatic abilities of the KKU-213L5 cells were compared with the parental line in vitro and in vivo. The expression profile of this metastatic cell line was determined using real-time PCR. KKU-213L5 cells were found to possess higher metastatic phenotypes, i.e., growth rates, stem cell surface markers (CD133), migration and invasion characteristics when compared with the parental cells. Compared to the KKU-213 cells, KKU-213L5 cells formed larger tumors in subcutaneous xenografted mice and had a >10-fold increase in lung metastases in the tail-vein injected metastatic mouse model. Mice injected intravenously with KKU-213L5 cells had a significantly shorter survival. Analysis of the expressed genes related to progression of cancer revealed significant upregulation of anterior gradient protein-2 (AGR2) and suppression of KiSS-1 in the KKU-213L5 cells. The association of these two genes with metastasis was affirmed in CCA patient tissues since increased AGR2 expression and decreased KiSS-1 expression were found in higher stage patient tumors. In conclusion, a highly metastatic human CCA cell line was established and characterized. It is plausible that the differential expression between the parental KKU-213 and highly metastatic KKU-213L5 cells may be beneficial to classify novel genes associated with metastasis. The KKU-213L5 cell line should serve as a valued device for discovering the molecular mechanisms of CCA metastasis and enabling the search for an effective therapy for the unmet clinical need in CCA.

Prevalence and hematological indicators of G6PD deficiency in malaria-infected patients.

BACKGROUND: This study aimed to evaluate the prevalence and alteration of hematological parameters in malaria patients with a glucose-6-phosphate dehydrogenase (G6PD) deficiency, in the western region of Thailand, an endemic region for malaria. METHODS: Data about patients with malaria hospitalized between 2013 and 2015 were collected. Clinical and sociodemographic characteristics such as age and gender, diagnosis on admission, and parasitological results were mined from medical records of the laboratory unit of the Phop Phra Hospital in Tak Province, Thailand. Venous blood samples were collected at the time of admission to hospital to determine G6PD deficiency by fluorescence spot test and detect malaria parasites by thick and thin film examination. Other data such as complete blood count and parasite density were also collected and analyzed. RESULTS: Among the 245 malaria cases, 28 (11.4 %) were diagnosed as Plasmodium falciparum infections and 217 cases (88.6 %) were diagnosed as P. vivax infections. Seventeen (6.9 %) patients had a G6PD deficiency and 228 (93.1 %) patients did not have a G6PD deficiency. Prevalence of male patients with G6PD deficiency was higher than that of female patients (P < 0.05, OR = 5.167). Among the patients with a G6PD deficiency, two (11.8 %) were infected with P. falciparum, while the remaining were infected with P. vivax. Malaria patients with a G6PD deficiency have higher monocyte counts (0.6 × 10(3)/µL) than those without a G6PD deficiency (0.33 × 10(3)/µL) (P < 0.05, OR = 5.167). Univariate and multivariate analyses also confirmed that malaria patients with a G6PD deficiency have high monocyte counts. The association between G6PD status and monocyte counts was independent of age, gender, nationality, Plasmodium species, and parasite density (P < 0.005). CONCLUSION: This study showed a prevalence of G6PD deficiency in a malaria-endemic area. This study also supported the assertion that patients with G6PD-deficient red blood cells had no protection against the P. falciparum infection. In addition, malaria patients with a G6PD deficiency had higher monocyte counts than those without a G6PD deficiency. These findings will help to recognize and diagnose malaria patients with a G6PD deficiency, as well as to identify the risks and protective factors against malaria in endemic areas.

A diagnostic tool for malaria based on computer software.

Nowadays, the gold standard method for malaria diagnosis is a staining of thick and thin blood film examined by expert laboratorists. It requires well-trained laboratorists, which is a time consuming task, and is un-automated protocol. For this study, Maladiag Software was developed to predict malaria infection in suspected malaria patients. The demographic data of patients, examination for malaria parasites, and complete blood count (CBC) profiles were analyzed. Binary logistic regression was used to create the equation for the malaria diagnosis. The diagnostic parameters of the equation were tested on 4,985 samples (703 infected and 4,282 control samples). The equation indicated 81.2% sensitivity and 80.3% specificity for predicting infection of malaria. The positive likelihood and negative likelihood ratio were 4.12 (95% CI = 4.01-4.23) and 0.23 (95% CI = 0.22-0.25), respectively. This parameter also had odds ratios (P value < 0.0001, OR = 17.6, 95% CI = 16.0-19.3). The equation can predict malaria infection after adjust for age, gender, nationality, monocyte (%), platelet count, neutrophil (%), lymphocyte (%), and the RBC count of patients. The diagnostic accuracy was 0.877 (Area under curve, AUC) (95% CI = 0.871-0.883). The system, when used in combination with other clinical and microscopy methods, might improve malaria diagnoses and enhance prompt treatment.

Cepharanthine suppresses metastatic potential of human cholangiocarcinoma cell lines.

Cholangiocarcinoma (CCA) is a highly metastatic tumor with poor responses to traditional chemotherapeutic agents. We have focused on new drugs which can exert effects either alone or in combination with available agents for a better treatment of CCA and recently reported efficacy of cepharanthine (CEP), a natural biscoclaurine alkaloid extract with anti-proliferative activity against human CCA cell lines. CEP treatment effectively suppressed tumor growth in CCA-inoculated mice without serious side-effects and also increased cell apoptosis in primary histocultures of CCA patient tissues, suggesting therapeutic potential of CEP against human CCA. In the present study, we further showed anti-metastatic effects of CEP on migration and invasion of human CCA cell lines. CEP at 10 µg/mL effectively suppressed migration activity of KKU-M213 and KKU-M214 as shown by wound healing and Boyden chamber assays. Similar suppressive effects were observed in invasion assays. The molecular mechanism underlying CEP actions on cell motility and invasion could be shown to involve suppression of ICAM-1 and MMP-2. Our results suggest CEP as a potential therapeutic agent for treating metastatic CCA.