Breath biopsy of breast cancer using sensor array signals and machine learning analysis.

Breast cancer causes metabolic alteration, and volatile metabolites in the breath of patients may be used to diagnose breast cancer. The objective of this study was to develop a new breath test for breast cancer by analyzing volatile metabolites in the exhaled breath. We collected alveolar air from breast cancer patients and non-cancer controls and analyzed the volatile metabolites with an electronic nose composed of 32 carbon nanotubes sensors. We used machine learning techniques to build prediction models for breast cancer and its molecular phenotyping. Between July 2016 and June 2018, we enrolled a total of 899 subjects. Using the random forest model, the prediction accuracy of breast cancer in the test set was 91% (95% CI: 0.85-0.95), sensitivity was 86%, specificity was 97%, positive predictive value was 97%, negative predictive value was 97%, the area under the receiver operating curve was 0.99 (95% CI: 0.99-1.00), and the kappa value was 0.83. The leave-one-out cross-validated discrimination accuracy and reliability of molecular phenotyping of breast cancer were 88.5 ± 12.1% and 0.77 ± 0.23, respectively. Breath tests with electronic noses can be applied intraoperatively to discriminate breast cancer and molecular subtype and support the medical staff to choose the best therapeutic decision.

A novel lung alveolar cell model for exploring volatile biomarkers of particle-induced lung injury.

Quartz can increase oxidative stress, lipid peroxidation, and inflammation. The objective of this study was to explore the volatile biomarkers of quartz-induced lung injury using a lung alveolar cell model. We exposed the human alveolar A549 cell line to 0, 200, and 500 µg/mL quartz particles for 24 h and used gas chromatography-mass spectrometry to measure the volatile metabolites in the headspace air of cells. We identified ten volatile metabolites that had concentration-response relationships with particles exposure, including 1,2,4-oxadiazole, 5-(4-nitrophenyl)-3-phenyl- (CAS: 28825-12-9), 2,6-dimethyl-6-trifluoroacetoxyoctane (CAS: 61986-67-2), 3-buten-1-amine, N,N-dimethyl- (CAS: 55831-89-5), 2-propanol, 2-methyl- (CAS: 75-65-0), glycolaldehyde dimethyl acetal (CAS: 30934-97-5), propanoic acid, 2-oxo-, ethyl ester (CAS: 617-35-6), octane (CAS: 111-65-9), octane, 3,3-dimethyl- (CAS: 4110-44-5), heptane, 2,3-dimethyl- (CAS: 3074-71-3) and ethanedioic acid, bis(trimethylsilyl) ester (CAS: 18294-04-7). The volatile biomarkers are generated through the pathways of propanoate and nitrogen metabolism. The volatile biomarkers of the alkanes and methylated alkanes are related to oxidative and lipid peroxidation of the cell membrane. The lung alveolar cell model has the potential to explore the volatile biomarkers of particulate-induced lung injury.

Targeting a cysteine protease from a pathobiont alleviates experimental arthritis.

BACKGROUND: Several lines of evidence suggest that the pathobiont Porphyromonas gingivalis is involved in the development and/or progression of auto-inflammatory diseases. This bacterium produces cysteine proteases, such as gingipain RgpA, endowed with the potential to induce significant bone loss in model systems and in patients. OBJECTIVE: We sought to gain further insight into the role of this pathobiont in rheumatoid arthritis (RA) and to identify novel therapeutic targets for auto-inflammatory diseases. METHODS: We profiled the antibody response to RgPA-specific domains in patient sera. We also tested the potential protective effects of RgpA domains in an experimental arthritis model. RESULTS: Pre-immunization of rats with purified recombinant RgpA domains alleviated arthritis in the joints of the rodents and reduced bone erosion. Using a functional genomics approach at both the mRNA and protein levels, we report that the pre-immunizations reduced arthritis severity by impacting a matrix metalloprotease characteristic of articular injury, a chemokine known to be involved in recruiting inflammatory cells, and three inflammatory cytokines. Finally, we identified an amino acid motif in the RgpA catalytic domain of P. gingivalis that shares sequence homology with type II collagen. CONCLUSION: We conclude that pre-immunization against gingipain domains can reduce the severity of experimentally induced arthritis. We suggest that targeting gingipain domains by pre-immunization, or, possibly, by small-molecule inhibitors, could reduce the potential of P. gingivalis to translocate to remote tissues and instigate and/or exacerbate pathology in RA, but also in other chronic inflammatory diseases.

Phytochemical profile of hot water extract of Glechoma hederacea and its antioxidant, and anti-inflammatory activities.

Glechoma hederacea belongs to the Labiatae family and has many biological activities. The objective of this study was to evaluate the chemical composition, antioxidant and anti-inflammatory activities of a hot water extract of G. hederacea (HWG). Our results indicated that rosmarinic acid, chlorogenic acid, caffeic acid, rutin, genistin, and ferulic acid were the most abundant phytochemicals in HWG. The free radical scavenging capacity of HWG in cell-free systems was evaluated by using the α,α-diphenyl-ß-picrylhydrazyl (DPPH) and ß-carotene bleaching assays. The antioxidant and anti-inflammatory activities of HWG were determined in vitro in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The results demonstrated that DAPI staining, the comet assay, and DNA fragmentation showed that HWG prevented LPS-induced DNA damage in RAW264.7 macrophages, reduced the content of LPS-induced nitric oxide (NO) and malondialdehyde (MDA), increased GSH levels, and regulated antioxidant enzyme activities. We also demonstrated that HWG significantly decreased the LPS-induced mRNA expression of iNOS, COX-2, TNF-α, IL-6, and IL-1ß in RAW264.7 macrophages, and reduced the LPS-induced protein expression of iNOS and COX-2 in RAW264.7 macrophages. These results show that HWG and its main components possess potent antioxidant and anti-inflammatory properties.

Stomach Cancer and Exposure to Talc Powder without Asbestos via Chinese Herbal Medicine: A Population-Based Cohort Study.

The present investigation was designed to explore the risk of stomach cancer by oral intake of talc powder without asbestos. We conducted a population-based cohort study on a randomly sampled cohort from Taiwan's health insurance database, with population of 1,000,000. The study participants were followed up through 2013. The outcome event of interest was the diagnosis of stomach cancer. The exposure of interest was the prescription of talc powder. Cox regression analyses were performed respectively. There were 584,077 persons without talc exposure and 21,575 talc users, 1849 diagnosed with stomach cancer. Persons with exposure of talc had a higher hazard ratio of stomach cancer (adjusted hazard ratio, 2.13; 95% confidence interval (CI), 1.54⁻2.94; p < 0.001). Classification by cumulative exposure of talc yielded adjusted hazard ratios of stomach cancer of 1.58 (95% CI, 0.79⁻3.17; p = 0.19) and 2.30 (95% CI, 1.48⁻3.57; p < 0.001) among persons with high (>21 g) and medium (6⁻21 g) exposure of talc, as compared to the low-exposure counterparts. Our data demonstrated positive association between increased risk of stomach cancer and oral intake of talc without asbestos. Despite the absence of dose-response effect, there might be a link between stomach cancer and talc.

A Study of Diagnostic Accuracy Using a Chemical Sensor Array and a Machine Learning Technique to Detect Lung Cancer.

Lung cancer is the leading cause of cancer death around the world, and lung cancer screening remains challenging. This study aimed to develop a breath test for the detection of lung cancer using a chemical sensor array and a machine learning technique. We conducted a prospective study to enroll lung cancer cases and non-tumour controls between 2016 and 2018 and analysed alveolar air samples using carbon nanotube sensor arrays. A total of 117 cases and 199 controls were enrolled in the study of which 72 subjects were excluded due to having cancer at another site, benign lung tumours, metastatic lung cancer, carcinoma in situ, minimally invasive adenocarcinoma, received chemotherapy or other diseases. Subjects enrolled in 2016 and 2017 were used for the model derivation and internal validation. The model was externally validated in subjects recruited in 2018. The diagnostic accuracy was assessed using the pathological reports as the reference standard. In the external validation, the areas under the receiver operating characteristic curve (AUCs) were 0.91 (95% CI = 0.79⁻1.00) by linear discriminant analysis and 0.90 (95% CI = 0.80⁻0.99) by the supportive vector machine technique. The combination of the sensor array technique and machine learning can detect lung cancer with high accuracy.

Melaleuca quinquenervia essential oil inhibits α-melanocyte-stimulating hormone-induced melanin production and oxidative stress in B16 melanoma cells.

BACKGROUND: Essential oils are odorous, volatile products of plant secondary metabolism, which are found in many leaves and stems. They show important biological activities, which account for the development of aromatherapy used in complementary and alternative medicine. The essential oil extracted from Melaleuca quinquenervia (Cav.) S.T. Blake (paperbark) (MQ-EO) has various functional properties. PURPOSE: The aim of this study is to investigate the chemical composition of MQ-EO by using gas chromatography-mass spectrometry (GC-MS) and evaluate its tyrosinase inhibitory activity. METHODS: Gas chromatography-mass spectrometry (GC-MS)-based metabolomics was used to identify 18 components in MQ-EO. The main components identified were 1,8-cineole (21.60%), α-pinene (15.93%), viridiflorol (14.55%), and α-terpineol (13.73%). B16 melanoma cells were treated with α-melanocyte-stimulating hormone (α-MSH) in the presence of various concentrations of MQ-EO or its major compounds. Cell viability was accessed by MTT assay and cellular tyrosinase activity and melanin content were determined by using spectrophotographic methods. The antioxidant mechanism of MQ-EO in α-MSH stimulated B16 cells was also investigated. RESULTS: In α-melanocyte-stimulating hormone (α-MSH)-stimulated murine B16 melanoma cells, MQ-EO, 1,8-cineole, α-pinene, and α-terpineol significantly reduced melanin content and tyrosinase activity. Moreover, MQ-EO, 1,8-cineole, α-pinene, and α-terpineol decreased malondialdehyde (MDA) levels. In addition, restored glutathione (GSH) levels, glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase activities were increased in α-MSH-stimulated B16 cells. MQ-EO not only decreased apoptosis but also reduced DNA damage in α-MSH stimulated B16 cells. These results showed that MQ-EO and its main components, 1,8-cineole, α-pinene, and α-terpineol, possessed potent anti-tyrosinase and anti-melanogenic activities besides the antioxidant properties. CONCLUSIONS: The active functional components of MQ-EO were found to be 1,8-cineole, α-pinene, and α-terpineol. Consequently, the results of present study suggest that MQ-EO is non-cytotoxic and can be used as a skin-whitening agent, both medically and cosmetically.

Diagnostic accuracy of breath tests for pneumoconiosis using an electronic nose.

Breath analyses have attracted substantial attention as screens for occupational environmental lung disease. The objective of this study was to develop breath tests for pneumoconiosis by analysing volatile organic compounds using an electronic nose. A case-control study was designed. We screened 102 subjects from a cohort of stone workers. After excluding three subjects with poorly controlled diabetes mellitus and one subject with asthma, 98 subjects were enrolled, including 34 subjects with pneumoconiosis and 64 healthy controls. We analysed the subjects' breath using an electronic nose with 32 nanocomposite sensors. Data were randomly split into 80% for model building and 20% for validation. Using a linear discriminate analysis, the sensitivity, specificity, accuracy, and area under the receiver operating characteristic curve (AUROC) were 67.9%, 88.0%, 80.8%, and 0.91, respectively, in the training set and 66.7%, 71.4%, 70.0%, and 0.86, respectively, in the test set. In subgroup analysis divided by smoking status, the AUROCs for current smokers, former smokers, and subjects who never smoked were 0.94, 0.93, and 0.99, respectively. In subgroup analysis divided by gender, the AUROCs for males and females were 0.95 and 0.99, respectively. Breath tests may have potential as a screen for pneumoconiosis. A multi-centre study is warranted, and the procedures must be standardized before clinical application.

Chemical characterization and antioxidative activity of four 3-hydroxyl-3-methylglutaroyl (HMG)-substituted flavonoid glycosides from Graptopetalum paraguayense E. Walther.

BACKGROUND: Graptopetalum paraguayense E. Walther is a popular traditional Chinese herb and possesses several health benefits. In earlier studies, we demonstrated that G. paraguayense showed no genotoxicity and showed several biological activities. However, the constituents of G. paraguayense have not been studied yet. In this present study, we isolated and identified the constituents of the leaves of G. paraguayense E. Walther. RESULTS: A total of seven flavonoid compounds were isolated from the methanolic extract of G. paraguayense. The four major compounds isolated were flavonoid glucoside derivatives of quercetin (1, 3) and kampferol (2, 4), each presenting a 3-hydroxyl-3-methylglutaroyl (HMG) substituent; compounds 3 and 4-the 2´´-acetyl derivatives of 1 and 2, respectively-are novel compounds isolated from nature for the first time. High-performance liquid chromatography for the quantitative analyses of the four major HMG-substituted flavonoid glycosides in G. paraguayense E. Walther were accomplished to acquire the high yields of 1-4 in the methanolic extract (4.8, 5.7, 4.3, and 2.5 mg/g, respectively). Furthermore, the antioxidant activities, including radical-scavenging, reducing power and lipid peroxidation inhibitory effects of these isolated flavonoids were also evaluated. All seven of the isolated flavonoid compounds possessed antioxdative activity. CONCLUSIONS: In this study of the constituents of the leaves of G. paraguayense E. Walther, we isolated four major components from its methanolic extract and determined their structures to be (acetylated) HMG-substituted flavonol glycosides, which are rare in nature. All seven of the isolated compounds possessed antioxdative activity, and those flavonoid compounds may be responsible for the functional ingredients in G. paraguayense. Further investigation of their bioactivities or pharmacological activities will be continued.

The melanogenesis alteration effects of Achillea millefolium L. essential oil and linalyl acetate: involvement of oxidative stress and the JNK and ERK signaling pathways in melanoma cells.

The mitogen-activated protein kinase (MAPK) family, including extracellular signal-regulated kinase (ERK)1/2, c-Jun N-terminal kinase (JNK)1/2 and p38 MAPK, is known to be activated by ultraviolet (UV) radiation in melanocytes to regulate melanin production. Reactive oxygen species (ROS) play important roles in the pathway of ERK and JNK activation. It has been established that the essential oil of Achillea millefolium L. (AM-EO) has activities that suppress the oxidative stress and inflammatory responses. Thus, we analyzed the effects of AM-EO on melanogenesis in melanocyte stimulating hormone (α-MSH) treated melanoma cells. The results demonstrated that AM-EO suppresses melanin production by decreasing tyrosinase activity through the regulation of the JNK and ERK signaling pathways. This effect might be associated with the AM-EO activity leading to the suppression of ROS, and linalyl acetate is its major functional component. Therefore, we propose that AM-EO has the potential to treat hyperpigmentation in the future.

Effect of Vetiveria zizanioides essential oil on melanogenesis in melanoma cells: downregulation of tyrosinase expression and suppression of oxidative stress.

The major objective of this study was to estimate the hypopigmentation function of the essential oil from Vetiveria zizanioides (VZ-EO). Our results indicated that VZ-EO exhibits potent lipid peroxidation inhibitory activity to moderate the bleaching of ß-carotene and to maintain the cellular glutathione (GSH) levels. VZ-EO can markedly decrease melanin production and tyrosinase activity in α-melanin-stimulating-hormone- (α-MSH-) stimulated B16 cells. The effect of VZ-EO on melanogenesis is achieved by the suppression of cellular tyrosinase expression. The results demonstrated that the activity of VZ-EO on melanogenesis might be the result of its potent antioxidative ability, which was reflected in the decreased cellular oxidant and malondialdehyde (MDA) levels and the recovered activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) in α-MSH-stimulated B16 cells. The most abundant compound in VZ-EO is cedr-8-en-13-ol (12.4%), which has a strong capability to inhibit lipid peroxidation. Therefore, VZ-EO has the potential to become an ingredient in future hypopigmentation drugs, foods, and cosmetics.

Achillea millefolium L. essential oil inhibits LPS-induced oxidative stress and nitric oxide production in RAW 264.7 Macrophages.

Achillea millefolium L. is a member of the Asteraceae family and has been used in folk medicine in many countries. In this study, 19 compounds in A. millefolium essential oil (AM-EO) have been identified; the major components are artemisia ketone (14.92%), camphor (11.64%), linalyl acetate (11.51%) and 1,8-cineole (10.15%). AM-EO can suppress the inflammatory responses of lipopolysaccharides (LPS)-stimulated RAW 264.7 macrophages, including decreased levels of cellular nitric oxide (NO) and superoxide anion production, lipid peroxidation and glutathione (GSH) concentration. This antioxidant activity is not a result of increased superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities, but rather occurs as a result of the down-regulation of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and heme oxygenase-1 (HO-1) expression, thus reducing the inflammatory response. Therefore, AM-EO can be utilized in many applications, including the treatment of inflammatory diseases in the future.

Improving antioxidant status in aged mice by 50% ethanol extract from red bean fermented by Bacillus subtilis.

BACKGROUND: The purpose of this study was to investigate the effects of 50% ethanol extracts from red bean non-fermented (RBE) and fermented by Bacillus subtilis (RBNE) on the antioxidant status of aged ICR mouse. RESULTS: Compared to 2-month-old ICR mouse, the plasma total antioxidant status (TAS) in 12-month-old ICR mouse decreased about 57%, while malondialdehyde (MDA) levels in the liver and brain of 12-month-old ICR mouse increased 56% and 30%, respectively. Orally administration of RBE or RBNE could completely recover the changes of MDA and plasma TAS levels due to the aging process. Vitamin E contents declined 88% in the liver and 74% in the brain of aged ICR mouse. At a level of 0.3 or 0.6 g kg(-1) body weight, RBNE raised vitamin E content in the liver and brain; however, RBE showed no significant influence. All antioxidant enzymes activities in the liver and brain of aged ICR mouse decreased compared to those activities in 2-month-old ICR mouse. RBNE could significantly enhance the superoxide dismutase activity in the brain of aged ICR mouse. CONCLUSION: Oral administration of RBE or RBNE could improve antioxidant status in aged ICR mouse. Fermentation by Bacillus subtilis could enhance the antioxidant properties of red bean.

Modifications of the defense and remodeling functionalities of bovine neutrophils inside the mammary gland of milk stasis cows received a commercial dry-cow treatment.

An appropriate length of milk stasis between two consecutive lactations of dairy cows is crucial for sustainable milk production. This dry period of cows allows extensive remodeling and sufficient cell renewal in mammary gland. Nevertheless, early dry period is one of the most risky stages in cow lactation cycle to intramammary infection. Dry-cow treatment through teats is, therefore, widely practiced at the commencement of milk stasis. Neutrophils are the most abundant cellular components in cow mammary secretion during early dry period, which in turn attribute to the meanwhile elevation of somatic cell counts and matrix metalloproteinase-9 (MMP-9) level. This study used bovine peripheral neutrophils as a cell model to examine the mode of modifications in their defense and remodeling functionalities after infiltration into mammary gland during early dry period. Results indicate a dose-dependent suppression of phorbol 12-myristate 13-acetate-stimulated free radical production and induction of MMP 9 degranulation in bovine peripheral neutrophils exposure to the d 7-dry secretion of cows received dry cow treatment at d 0 in milk stasis. Meanwhile, an enhancement of plasminogen activation and TNF-α shedding on bovine peripheral neutrophils were also observed. These two cellular events might be involved in the functional modifications on infiltrated neutrophils during early dry period. In conclusion, the opposite trend of modifications in the defense and matrix remodeling functionalities of neutrophils inside the mammary gland of cows at early dry period reflect the collaboration of infiltrated neutrophils for promoting extensive glandular remodeling at minimum compromise of local defense during the acute involution period without apparent disturbance by dry cow treatment.

Zanthoxylum ailanthoides Sieb and Zucc. extract inhibits growth and induces cell death through G2/M-phase arrest and activation of apoptotic signals in colo 205 human colon adenocarcinoma cells.

The effects of 50% ethanolic stem extracts of Zanthoxylum ailanthoides Sieb and Zucc. (ZASZ) on the cell viability, cell cycle and apoptosis were investigated in a human colon adenocarcinoma cell line (colo 205). The results demonstrated that ZASZ induced morphological changes and decreased the cell viability. ZASZ promoted Wee1, checkpoint kinase 2 (CHK2), p21 and p53 levels, decreased cyclin B and cdc25c associated with that led to G(2)/M phase arrest. ZASZ-triggered apoptosis was confirmed by 4' -6-diamidino-2-phenylindole (DAPI) staining and DNA gel electrophoresis. ZASZ increased the levels of glucose-regulated protein 78 (GRP78) and growth arrest and DNA damage inducible gene 153 (GADD153), and promoted an increase of reactive oxygen species (ROS) and Ca(2+) release, and loss of mitochondrial membrane potential (ΔΨ(m)) accompanied by cytochrome c release that was due to the decrease of Bcl-2 and increase of Bax levels in the colo 205 cells. ZASZ also induced the protein levels of apoptosis-inducing factor (AIF) and endonuclease G (Endo G), increased the levels of caspase-3, -7 and -9, and stimulated the levels of fatty acid synthase (Fas) and Fas ligand in the colo 205 cells. ZASZ contains phenolic compounds, including flavone, chlorogenic acid and isofraxidin, among which, flavone was found to be the most effective in reducing cell viability and proliferative responses in the colo 205 cells. ZASZ induces cytotoxicity and apoptosis in colo 205 cells which provides the rationale for studies in animal models on the utilization of ZASZ as a potential cancer therapeutic compound.

Isolation of substances with antiproliferative and apoptosis-inducing activities against leukemia cells from the leaves of Zanthoxylum ailanthoides Sieb. & Zucc.

Extraction of the leaves of Zanthoxylum ailanthoides Sieb. & Zucc. affords extracts and four isolated compounds which exhibit activities against leukemia cells. The chloroform-soluble fraction (ZAC) of the crude extract of this plant showed cytotoxic activity against human promyelocytic leukemia (HL-60) and myelomonocytic leukemia (WEHI-3) cells with IC(50) values of 73.06 and 42.22 µg/mL, respectively. The active ZAC was further separated to yield pheophorbide-a methyl ester (1), pheophorbide-b methyl ester (2), 13(2)-hydroxyl (13(2)-S) pheophorbide-a methyl ester (3) and 13(2)-hydroxyl (13(2)-R) pheophorbide-b methyl ester (4) whose structures were confirmed by spectroscopic methods. Compounds 2-4 showed cytotoxic activities against both leukemia cells with IC(50) value in the range of 46.76-79.43 nM, whereas compound 1 exhibited only weak cytotoxic activity. The extracts and compounds 1-4 also induced apoptosis and DNA damage in leukemia cells after treatment. The results suggested that the Z. ailanthoides is biologically active against leukemia cells.

Hepatoprotection of chlorella against carbon tetrachloride-induced oxidative damage in rats.

The effects of 80% ethanolic chlorella extracts (GPE) on carbon tetrachloride (CCl(4))-induced hepatic damage were investigated in Sprague-Dawley rats. The rats were orally treated with GPE (0.5 g/kg body weight) or silymarin (0.2 g/kg body weight) over four consecutive weeks with administration of CCl(4) (20% CCl(4), 0.5 ml/rat twice a week). The GPE had a significant protective effect against liver injuries, as well as oxidative stress induced by CCl(4), resulting in reduced lipid peroxidation and improved serum biochemical parameters such as aspartate aminotransferase and alanine aminotransferase. The reduced levels of glutathione, vitamin C, superoxide dismutase, and catalase in the CCl(4)-treated rats were significantly increased by treatment with GPE. Furthermore, the activity of GPE was comparable to the standard drug silymarin. In conclusion, chlorella may be useful as a hepatoprotective agent against chemical-induced liver damage in vivo.

Association between Interleukin-10 Gene Promoter Haplotype and Schizophrenia in a Han-Chinese Study.

Schizophrenia is a multi-factorial genetic disease, and it is caused by a combination of different gene polymorphisms and not individual ones, however, its pathogenesis is still unclear. The purpose of this study was explored the association between the -1082G/A, -819T/C, and -592C/A polymorphisms of interleukin-10 (IL-10) in schizophrenia. A total of 659 schizophrenics were recruited from a teaching hospital, whereas 411 healthy non-schizophrenic individuals were recruited from community in the same geographical area. The -1082G/A, -819T/C and -592C/A polymorphisms were genotyped by using PCR-RFLP, direct sequencing and TaqMan(®) SNP assay. Both maximum likelihood method of UNPHASED program and Bayesian method of PHASE software were utilized for haplotypic analysis. An allelic frequency difference was found between the schizophrenics and community controls at -1082G/A polymorphism of IL-10 promoter (χ(2) =4.678, P=0.031). A haplotype of ACA was observed to be associated with schizophrenia after performing UNPHASED, PHASE and multivariate logistic regression analysis (P<0.001; P=0.001). In addition, the persons who carry haplotype ACA of IL-10 promoter SNPs were estimated for 5.789 fold higher risk to develop schizophrenia than controls. We postulated this haplotype association might due to variant-specific effect on IL-10 gene regulation, which leads to imbalance secretion of Th1/Th2 cytokines. Nevertheless, more detailed mechanism needs to be elucidated in further investigations in order to confirm this hypothesis.