Search

Background Danshen is a common traditional Chinese medicine used to treat neoplastic and chronic inflammatory diseases in China. However, the effects of Danshen on human oral cancer cells remain relatively unknown. This study investigated the antiproliferative effects of a Danshen extract on human oral cancer SAS, SCC25, OEC-M1, and KB drug-resistant cell lines and elucidated the possible underlying mechanism. Methods We investigated the anticancer potential of the Danshen extract in human oral cancer cell lines and an in vivo oral cancer xenograft mouse model. The expression of apoptosis-related molecules was evaluated through Western blotting, and the concentration of in vivo apoptotic markers was measured using immunohistochemical staining. The antitumor effects of 5-fluorouracil and the Danshen extract were compared. Results Cell proliferation assays revealed that the Danshen extract strongly inhibited oral cancer cell proliferation. Cell morphology studies revealed that the Danshen extract inhibited the growth of SAS, SCC25, and OEC-M1 cells by inducing apoptosis. The Flow cytometric analysis indicated that the Danshen extract induced cell cycle G0/G1 arrest. Immunoblotting analysis for the expression of active caspase-3 and X-linked inhibitor of apoptosis protein indicated that Danshen extract-induced apoptosis in human oral cancer SAS cells was mediated through the caspase pathway. Moreover, the Danshen extract significantly inhibited growth in the SAS xenograft mouse model. Furthermore, the Danshen extract circumvented drug resistance in KB drug-resistant oral cancer cells. Conclusion The study results suggest that the Danshen extract could be a potential anticancer agent in oral cancer treatment.

// Cheng-Yu Yang 1, 6 , Chih-Kung Lin 3 , Chang-Huei Tsao 7, 8 , Cheng-Chih Hsieh 5 , Gu-Jiun Lin 4 , Kuo-Hsing Ma 4 , Yi-Shing Shieh 6 , Huey-Kang Sytwu 1, 7 and Yuan-Wu Chen 1, 2, 6 1 Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan 2 Department of Oral and Maxillofacial Surgery, Tri-Service General Hospital, Taipei, Taiwan 3 Division of Anatomic Pathology, Taipei Tzu Chi Hospital, Taipei, Taiwan 4 Department of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan 5 Department of Pharmacy Practice, Tri-Service General Hospital, Taipei, Taiwan 6 School of Dentistry, National Defense Medical Center, Taipei, Taiwan 7 Graduate Institute of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan 8 Department of Medical Research, Tri-Service General Hospital, Taipei, Taiwan Correspondence to: Yuan-Wu Chen, email: h6183@yahoo.com.tw Keywords: patient-derived tumor xenograft, LSD1, melatonin, oral cancer, cell proliferation Received: June 16, 2016 Accepted: March 08, 2017 Published: April 04, 2017 ABSTRACT Aberrant activation of histone lysine-specific demethylase (LSD1) increases tumorigenicity; hence, LSD1 is considered a therapeutic target for various human cancers. Although melatonin, an endogenously produced molecule, may defend against various cancers, the precise mechanism involved in its anti-oral cancer effect remains unclear. Patient-derived tumor xenograft (PDTX) models are preclinical models that can more accurately reflect human tumor biology compared with cell line xenograft models. Here, we evaluated the anticancer activity of melatonin by using LSD1-overexpressing oral cancer PDTX models. By assessing oral squamous cell carcinoma (OSCC) tissue arrays through immunohistochemistry, we examined whether aberrant LSD1 overexpression in OSCC is associated with poor prognosis. We also evaluated the action mechanism of melatonin against OSCC with lymphatic metastases by using the PDTX models. Our results indicated that melatonin, at pharmacological concentrations, significantly suppresses cell proliferation in a dose- and time-dependent manner. The observed suppression of proliferation was accompanied by the melatonin-mediated inhibition of LSD1 in oral cancer PDTXs and oral cancer cell lines. In conclusion, we determined that the beneficial effects of melatonin in reducing oral cancer cell proliferation are associated with reduced LSD1 expression in vivo and in vitro .

Objectives This study aimed to evaluate the adverse clinicopathologic features of oral squamous cell carcinoma (OSCC), including margin status, depth of invasion, lymphovascular invasion, perineural invasion, and extranodal extension that significantly affect survival outcomes. Materials and methods This retrospective cross-sectional study included 341 patients with OSCC who underwent therapeutic surgical treatment in Taiwan. The Kaplan–Meier method was used to estimate survival outcomes. A multivariable Cox regression model was used to evaluate the associations of various clinicopathologic features with 5-year overall survival (OS) outcomes in patients with pN0 and pN+ tumors. Results Overall, the patients had 5-year OS and progression-free survival rates of 60.0 and 47.9%, respectively. In the pN0 group, the multivariate analysis identified a positive margin (odds ratio [OR] = 16.3, 95% confidence interval [95% CI]: 3.7–72.3; P = 0.001), depth of invasion >5 mm (OR = 2.1, 95% CI: 1.2–3.7; P = 0.012), presence of lymphovascular space invasion (OR = 5.4, 95% CI: 1.3–22.0; P = 0.018), and presence of perineural invasion (OR = 4.3, 95% CI: 1.7–11.1; P = 0.002) as independent and significant prognosticators of OS. In the pN+ group, only the presence of extranodal extension independently predicted OS (OR = 1.7, 95% CI: 1.1–2.7; P = 0.0026). Conclusions When determining survival prognosis for patients with a pN0 status, we recommended including all adverse features. In contrast, extranodal extension was the most important prognostic factor for patients with a pN+ status.

Background: Biological and prognostic roles of programmed death ligand-1 (PD-L1) remain unclear in oral squamous cell carcinoma (OSCC). The controversies role of tumor microenvironmental IFN-γ, pivotal roles in host responses to malignant cells, in oral cancer growth and PD-L1 expression was under-investigated. We examined the mechanism of PD-L1 regulation with focus on interferon gamma in vitro and in clinical samples. Triptolide is the bioactive compound isolated from Tripterygium wilfordii, which has been reported exhibit anti‑inflammatory and antitumor activities. Moreover, we investigated whether TPL affected suppression of PD-L1 is involved in its anti-cancer activity. Methods: The PD-L1 expressions were analyzed by immunohistochemistry in 135 tissues samples. We analysed the production of IFN-γ at oral cancer cell by enzyme-linked immunosorbent assay (ELISA) analysis. IFN-ϒ on oral cancer cell proliferation were examined. We investigated the anticancer of the TPL on human oral cancer by targeted PD-L1 in vitro and patient-derived tumor xenograft (PDTX) models. Findings: PD-L1 overexpression was significant in the tumor site of oral cancer (P < .01). IFN-γ driven the PD-L1 expression oral cancer cells in a dose-dependent manner. The TPL inhibits tumor growth in oral cancer PDTX model with suppressing the PD-L1 expression. The TPL also suppresses the oral cancer cell PD-L1 expression in an interferon-gamma microenvironment. Interpretation: Our results provide evidence for clinical development of PD-L1 targeting therapy in OSCC treatment in future. Funding Statement: This study was supported by research grants from Tri-Service General Hospital, Taiwan, Republic of China (grants Nos. TSGH-C106-004- 006-008-S05, TSGH-C107-008-S06, TSGH-C108-007-008-S06), Ministry of Science and Technology, Taiwan, Republic of China (grants No. MOST 105- 2314-MAB-106-090). Declaration of Interests: All authors have completed the disclosure forms and declare that: no support, financial or otherwise, has been received from any organization that may have an interest in the submitted work; and there are no other relationships or activities that could appear to have influenced the submitted work. Ethics Approval Statement: The experiments were conducted according to the ethical guidelines of the institutional review board (TSGH-2-102-05-111) of the National Defense Medical Center, Taiwan.

Screening for oral potentially malignant disorders (OPMDs) with dysplasia in high-risk groups is suggested in countries with a high prevalence of the disorders. This study aimed to compare the accuracy of diagnoses of OPMDs with dysplasia made by a primary examiner (general dental clinician) and a specialist (oral and maxillofacial surgeon) using the current Taiwanese Nationwide Oral Mucosal Screening Program (TNOMSP). A total of 134 high-risk participants were enrolled for oral mucosal screening via the TNOMSP. A primary examiner and a specialist examined each participant. Mucosal biopsies were obtained and subjected to histopathological analysis. The OPMD most frequently diagnosed by the primary examiner was thin homogeneous leukoplakia (48/134; 35.8%), and in 39/134 participants (29.1%) the diagnosis was uncertain, but abnormalities were suggested. The OPMDs most frequently diagnosed by the specialist were erythroleukoplakia (23/134; 17.2%) and thin homogeneous leukoplakia (21/134; 15.7%), and 51/134 participants (38.1%) were diagnosed with other diseases. Via histopathology, 70/134 participants (52.3%) were diagnosed with dysplasia, and 58/134 (43.3%) were diagnosed with benign conditions. The specialist’s diagnoses exhibited a higher specificity, positive predictive value, and accuracy than the primary examiners. A specialist using the current TNOMSP for high-risk participants diagnosed OPMDs with dysplasia more accurately than a primary examiner. Early diagnosis of high-risk OPMDs is crucial in countries with a high prevalence of the disorders. Proficient examination via the current TNOMSP by trained clinician is effective for the management of OPMDs with dysplasia.

Biological and prognostic roles of programmed death ligand 1 (PD-L1) remain unclear in oral squamous cell carcinoma (OSCC). Moreover, the pivotal role of tumor microenvironmental interferon-gamma (IFN-γ) in host responses to malignant cells, oral cancer growth, and PD-L1 expression has not been adequately studied. Thus, PD-L1 expression in 130 OSCC samples was analyzed using immunohistochemistry, which was found significantly overexpressed at the tumor site (P

Background: Oral squamous cell carcinoma (OSCC) is the most frequently found malignant tumor of the oral cavity, and the annual death rate of OSCC patients has been increasing rapidly among the Taiwanese population. Aim: The aim of this study is to evaluate if poor nutrition is a potential adverse prognostic factor in patients with OSCC. Methods: This cross-sectional study included 322 patients with OSCC who underwent therapeutic surgical treatment in Taiwan. The pretreatment body mass index (BMI) and serum uric acid and albumin levels were measured as common indicators of the nutritional status, and overall survival (OS) was analyzed using the Kaplan–Meier method and a Cox regression model. Results: The optimal cut-off values were 24.0 kg/m[2], 5.05 mg/dL, and 3.85 g/dL for BMI, serum uric acid, and serum albumin, respectively. The multivariate Cox regression analysis indicated lower BMI (hazard ratio [HR] = 1.557, 95% confidence interval [95% CI]: 1.070–2.265; P = 0.021), lower serum uric acid (HR = 1.857, 95% CI: 1.265–2.724; P = 0.002), and lower serum albumin (HR = 2.011, 95% CI: 1.278–3.163; P = 0.003) as independent and significant prognosticators of OS. Advanced stage, depth of invasion of more than 5 mm, and extranodal extension were prognostic predictors of poor OS. Conclusion: A lower preoperative BMI and lower serum uric acid and albumin levels are predictors of significantly worse survival among OSCC patients. Maintaining a healthy nutritional status is crucial to improving the survival of patients with oral cancer. The treatment for patients with poor nutritional status should be more aggressive and cautious.

Dysregulation of histone demethylase Jumonji-C domain-containing protein 5 (JMJD5) has been identified as a great effect on tumorigenesis. Silibinin is a commonly used anti-hepatotoxic drug and exhibits anticancer effect in various cancers. However, the antitumor mechanism between silibinin and JMJD5 in oral squamous cell carcinoma (OSCC) remains unclear. In this study, the clinical significance of JMJD5 on OSCC patients was assessed through tissue microarray. Furthermore, mice bearing patient-derived tumor xenografts (PDTXs) and tongue cancer cell lines were treated with silibinin and evaluated for tumor growth and JMJD5 expression. High expression of JMJD5 in oral cancer was significantly associated with tumor size (P = 0.0241), cervical node metastasis (P = 0.0001) and clinical stage (P = 0.0002), was associated with worse survival rate compared with that of the total cohort (P = 0.0002). Collectively the data indicate that JMJD5 expression may be suitable for detection of unfavorable prognosis in OSCC patients, based in part on its apparent role as a marker of metastasis. In addition, silibinin inhibits cancer growth in vitro and in PDTX models. Furthermore, metastasis-associated protein 1 (MTA1) could regulate the expression for JMJD5 and had a positive correlation with JMJD5. Moreover, silibinin could downregulate JMJD5 and MTA1 in oral cancer. Present study thus identifies that JMJD5 might be an essential prognostic indicator and therapeutic target against OSCC progression. In addition, silibinin is a potential candidate among novel chemotherapeutic agents or adjuvants for modulating JMJD5 in OSCC, through a mechanism likely involving MTA1/JMJD5 axis.

The primary strategy for treating oral squamous cell carcinoma (OSCC) is therapeutic resection, with the trismus resection defect reconstructed via free flap. The most popular free flaps include the radial forearm free flap (RFFF) and anterolateral thigh free flap (ALT). This study investigated the relationships between the hospitalization period and a variety of surgical outcomes, as well as maximum inter-incisor distance (IID), in trismus patients who chewed betel nuts. Forty-nine primary OSCC patients who chewed betel nuts and underwent surgical resection and reconstruction between 2010 and 2016 were enrolled in this retrospective study. The data were from a single center in Taiwan. The outcome variable after flap recovery surgery was the duration of postoperative hospitalization. Other factors that were analyzed comprised correlations between hospitalization and a variety of factors, including postoperative inter-incisor distances (IIDs), operative time, gender, and WBC count, upon stratification into two reconstruction groups. The mean postoperative hospitalization duration in the ALT group was 22.9 ± 7.2 days, which was significantly shorter than that in the RFFF group (27.8 ± 7.0 days; p = 0.019). Two-week postoperative IID (ALT group: 16.1 ± 0.8 mm; RFFF group: 7.0 ± 0.6 mm) was inversely related to the duration of hospitalization (p = 0.022, r = − 0.372). The ALT flap is more effective than the RFFF flap to reduce the length of hospitalization in trismus patients. The ALT flap should be considered as a first-line technique in OSCC reconstruction in trismus patient reconstruction.

Background Aberrant expression of decoy receptor 3 (DcR3) is considered to be a diagnostic and therapeutic target for human cancers. The aim of this study was to assess DcR3 as a target of the anticancer effects of triptolide (TPL) in preclinical patient‐derived tumor xenograft (PDTX) models of oral squamous cell carcinoma (OSCC). Methods The expression of DcR3 was evaluated through immunohistochemistry, and correlations were examined using clinical variables. The effects of TPL on the expression of DcR3 and cell proliferation were investigated in OSCC cell lines and in PDTX models. Results DcR3 overexpression was associated with overall survival and tumor size. TPL significantly decreased tumor growth. Moreover, TPL inhibited the expression of metastasis‐associated protein 1 (MTA1), a transcription factor for DcR3 in vivo, in vitro, and in PDTX models. Conclusion TPL appeared to exert anticancer effects by repressing DcR3 and MTA1 in vitro, in vivo, and in PDTX models.

Results of previous studies demonstrated that the tetraindole, SK228, which has a high lipid but low water solubility, displayed moderate anticancer efficacy in a xenograft model of breast cancer. This finding led to the proposal that new, pyridine based tetraindole (PBT) analogs of SK228, containing tetraindole moieties distributed about central protonated pyridine cores, would have enhanced bioavailabilities and anticancer efficacies. Among the PBTs prepared and subjected to biological studies, 3f (FCW81) was observed to display the highest antiproliferative activity against the two triple negative breast cancer (TNBCs) cell lines, MDA-MB-231 and BT549. In addition, its mode of action was shown to involve G2/M arrest of the cell cycle along with the promotion of increased levels of cyclin B1 and p-chk2 and a decreased level of p-cdc2. DNA damage and induction of apoptosis caused by FCW81 was found to be associated with a decrease in DNA repair. Significantly, FCW81 displays therapeutic efficacy in a xenograft model of human breast cancer by not only serving to inhibit markedly the growth of cancer cells but also to block effectively cancer cell metastasis. Collectively, the results of these studies have led to the identification of novel pyridine-tetraindole based anticancer agents with potential use in TNBC therapy.

Objectives To evaluate whether low body mass index (BMI) is a potential adverse prognostic factor in patients with oral squamous cell carcinoma (OSCC). Material and methods This cross-sectional study included 320 patients with OSCC who underwent therapeutic surgical treatment in Taiwan. The pretreatment BMI was measured as a common indicator of the pretreatment nutritional status to calculate the overall survival in Kaplan-Meier method. The adverse histopathological features of margin status, depth of invasion (DOI), lymphovascular invasion (LVSI), perineural invasion (PNI), and extranodal extension (ENE) were analyzed using the Cox regression model. Results Low BMI (underweight), DOI > 5 mm, and ENE were identified as detrimental prognostic factors. On multivariate Cox regression analysis, the low BMI group (odds ratio [OR] = 1.683; 95% confidence interval [95% CI] 1.116–2.539; P = 0.022), DOI > 5 mm (OR = 2.399; 95% CI 1.459–3.943; P = 0.001), and ENE (OR = 2.467; 95% CI 1.540–3.951; P = 0.000) yielded reduced survival rate. Conclusions The lower BMI had an important and significant effect on the survival of patients with oral cancer and their surgical outcomes. In addition to the adverse histopathological features, a DOI > 5 mm and positive ENE were also identified as the most important prognostic factors. Clinical relevance Underweight patients with low BMI, DOI of > 5 mm, and positive ENE should receive more intensive nutritional supplementation and postoperative adjuvant therapy.

Objectives Lymph node metastasis in oral squamous cell carcinoma (OSCC) is a poor prognostic factor. The histopathologic stage (e.g., pN) is used to evaluate the severity of lymph node metastasis; however, the current staging system insufficiently predicts survival and recurrence. We investigated clinical outcomes and lymph node density (LND) in betel nut-chewing individuals. Material and methods We retrospectively analyzed 389 betel nut-exposed patients with primary OSCC who underwent surgical resection in 2002–2015. The prognostic significance of LND was evaluated by overall survival (OS) and disease-free survival (DFS) using the Kaplan-Meier method. Results Kaplan-Meier analyses showed that the 5-year OS and DFS rates in all patients were 60.9 and 48.9%, respectively. Multivariate analysis showed that variables independently prognostic for OS were aged population (hazard ratio [HR] = 1.6, 95% confidence interval [95% CI] = 1.1–2.5; P = .025), and cell differentiation classification (HR = 2.4, 95% CI = 1.4–4.2; P = .002). In pathologic N-positive patients, a receiver operating characteristic (ROC) curve for OS was used and indicated the best cutoff of 0.05, and the multivariate analysis showed that LND was an independent predictor of OS (HR = 2.2, 95% CI = 1.3–3.7; P = .004). Conclusions Lymph node density, at a cutoff of 0.05, was an independent predictor of OS and DFS. OS and DFS underwent multiple analyses, and LND remained significant. The pathologic N stage had no influence in the OS analysis. Clinical relevance LND is a more reliable predictor of survival in betel nut-chewing patients for further post operation adjuvant treatment, such as reoperation or adjuvant radiotherapy.

Mycobacterium tuberculosisinfection is a major world health issue. The early identification of patients at risk for a poor response to anti-M. tuberculosistherapy would help elucidate the key players in the anti-M. tuberculosisresponse. The objective of the present study was to correlate the modulation of cytokine expression (interleukin-1 [IL-1], IL-6, IL-8, IL-10, IL-12, gamma interferon [IFN-γ], interferon-inducible protein [IP-10], and monocyte chemotactic protein 1 [MCP-1]) with the clinical response to 2 months of intensive therapy. From January to December 2007, 40M. tuberculosis-infected patients and 40 healthy patients were recruited. After exclusion for diabetes, 32 patients and 36 controls were analyzed. The clinical responses of theM. tuberculosis-infected patients on the basis of the findings of chest radiography were compared to their plasma cytokine levels measured before and after 2 months of intensive anti-M. tuberculosistherapy and 6 months of therapy with human cytokine antibody arrays. Chest radiographs of 20 of 32M. tuberculosis-infected patients showed improvement after 2 months of intensive therapy (early responders), while theM. tuberculosisinfections in 12 of 32 of the patients resolved after a further 4 months (late responders). The levels of expression of TNF-α, MCP-1, IFN-γ, and IL-1β were decreased; and the level of IL-10 increased in early responders. After adjustment for age, gender, and the result of sputum culture forM. tuberculosis, significant differences in the levels of MCP-1 and IP-10 expression were observed between the early and the late responders after 2 months of intensive anti-M. tuberculosistherapy. Due to the interpatient variability in IP-10 levels, intrapatient monitoring of IP-10 levels may provide more insight into theM. tuberculosisresponder status than comparison between patients. Plasma MCP-1 levels were normalized in patients who had resolved theirM. tuberculosisinfections. Further studies to evaluate the association of the modulation in MCP-1 levels with early and late responses are warranted.

Objectives Advanced oral cancer is a major public health concern because of a lack of effective prevention and treatment. Triptolide (TPL), a diterpenoid triepoxide derived from the Chinese herb Tripterygium wilfordii, has been demonstrated to possess strong anticancer properties. In this study, we investigated whether TPL exerts anticancer effects on the tumor microenvironment of head and neck squamous cell carcinoma (HNSCC). Materials and methods Human macrophage-like U937 cells were co-inoculated with oral cancer SAS cells in a noncontact transwell coculture system. Cytokine expression was detected using ELISA, and cell proliferation was detected using methylene blue. RNA levels were detected using qPCR. Protein levels were detected using Western blot analysis. In vivo experiments involved using xenografted NOD/SCID mice. Results Our results demonstrated that TPL inhibited the growth of SAS cells co-inoculated with U937 cells in vitro and in vivo. TPL inhibited the invasion, migration ability, and angiogenesis of SAS cells co-inoculated with U937 cells. Expression of cytokines IL-6, IL-8, and TNF-α was induced by co-inoculation, but TPL repressed their expression. Conclusion TPL suppressed the expression of cytokines IL-6, IL-8, and TNF-α, as well as tumor growth, invasion, migration, and angiogenesis in the co-inoculation of human tongue cancer cells with macrophage-like U937 cells. Clinical relevance TPL is a potential candidate among novel chemotherapeutic agents or adjuvants for modulating tumor-associated macrophages in a tumor microenvironment of HNSCC.

Zinc oxide nanoparticles (ZnO NPs) are one of the most widely used nanomaterials in consumer products and industrial applications. As a result of all these uses, this has raised concerns regarding their potential toxicity. We previously found that candidate markers of idiopathic pulmonary fibrosis and lung cancer were significantly up-regulated in rat bronchoalveolar lavage fluid (BALF) following exposure to ZnO NPs by using a liquid chromatography (LC)-based proteomic approach. To achieve comprehensive protein identification analysis, we conducted the two-dimensional gel electrophosis (2-DE)-based proteomic workflow to analyze the differences in BALF proteins from rats that had been exposed to a high dose of 35 nm ZnO NPs. A total of 31 differentially expressed protein spots were excised from the gels and analyzed by nanoLC-tandem mass spectrometry (MS/MS). Gene ontology (GO) annotation of these proteins showed that most of the differentially expressed proteins were involved in response to stimulus and inflammatory response processes. Moreover, pulmonary surfactant-associated protein D and gelsolin, biomarkers of idiopathic pulmonary fibrosis, were significantly up-regulated in rat BALF after ZnO NPs exposure (2.42- and 2.84-fold, respectively). The results obtained from this present study could provide a complementary consequence with our previous study and contribute to a better understanding of the molecular mechanisms involved in ZnO NP-induced lung disorders.

Differentiating pulmonary tuberculosis (TB) from pneumonia caused by other infectious organisms remains a serious challenge. Examination of sputum smears for acid-fast bacilli (AFB) is relatively insensitive, and culture for Mycobacterium tuberculosis takes days to weeks to produce a result (10). The use of the tuberculin skin test (TST) to diagnose TB is limited in regions where a large fraction of the population has been vaccinated with Mycobacterium bovis bacillus Calmette-Guerin (BCG) (11). Therefore, recent research has focused on identifying M. tuberculosis-specific antigens. One of these, early secretory antigen target-6 (ESAT-6), is encoded by a genetic segment that is absent from BCG (25), and two commercial tests have been developed on the basis of the enhanced ability of ESAT-6 (and other antigens) to simulate production of gamma interferon (IFN-γ) in blood samples from patients with TB (18). Although useful for the diagnosis of TB, a study examining the utility of such tests for differentiating TB from other pulmonary diseases has demonstrated rather low specificity and sensitivity (76% and 79%, respectively), presumably because of the background of underlying respiratory disease in the patients (17). The objective of this study was to determine whether other cytokine markers could be identified that would be more useful for the differential diagnosis of TB and pneumonia from blood samples. We used microarrays coated with antibodies against tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), IL-8, IL-10, monocyte chemoattractant protein 1 (MCP-1), IFN-γ, IL-1β, IFN-induced protein 10 (IP-10), and IL-12 to evaluate the cytokine profiles of whole blood cells collected from patients with TB or pneumonia and from healthy individuals. The cytokine profiles were determined before and after stimulation with ESAT-6 (to magnify the cytokine response in patients with TB) and lipopolysaccharide ([LPS] to amplify the response in patients with pneumonia) in terms of both the absolute levels and the percent differences before and after stimulation.