Vitamin D levels in the assessment of Crohn's disease activity and their relation to nutritional status and inflammation.

BACKGROUND: Crohn's disease (CD) is frequently associated with malnutrition, inflammation and a deficiency of vitamin D (VD) with the relationships between these symptoms being poorly defined. VD is a modulator of the immune system and is associated with the onset of CD and disease activity. The level of serum VD may have potential in the assessment of CD activity. This study aimed to evaluate the relationships between VD, nutritional status and inflammation, and to identify more accurate VD thresholds. METHODS: The study included 76 outpatients with CD diagnosed between October 2018 and October 2020 and 76 healthy volunteers. Levels of serum 25(OH)D and nutritional indicators, as well as biochemical and disease activity assessments, were conducted. RESULTS: Patients with CD and healthy participants were found to differ significantly in their 25(OH)D levels as well in levels of nutritional and inflammatory indicators. The optimal VD cut-off value was found to be 46.81 nmol/L for CD development and 35.32 nmol/L for disease activity. Levels of 25(OH)D were correlated with both nutritional status and inflammation. CONCLUSIONS: The VD level is likely to be a useful additional tool in the evaluation of CD patients and predicting the disease activity and clinical response. The VD level may relate both to the nutritional status and levels of inflammation in CD patients, and disease progression.

Risk factors associated with catheter-related venous thrombosis: a meta-analysis.

BACKGROUND: So far, the risk factors of catheter-related venous thrombosis (CRVT) are not fully understood. We use evidence-based medicine to find the risk factors of CRVT by pooling the current studies that reported the risk factors of CRVT, aiming to provide guidance for clinical diagnosis and treatment. METHODS: We searched PubMed, Embase, and Cochrane Library from the establishment of the database to July 2021. We included studies that reported the risk factors of CRVT, and we excluded duplicate publications, research without full text, incomplete information or inability to conduct data extraction, animal experiments, reviews, and systematic reviews. STATA 15.1 was used to analyze the data. RESULTS: The pooled results show that history of venous thrombosis (odds ratio [OR] = 3.75, 95% confidence interval [CI]: 1.02-13.85; P = 0.047), cancer (OR = 1.74, 95% CI: 1.17-2.57; P = 0.006), infection (OR = 2.13, 95% CI:1.33-3.42; P = 0.002), and multilumina (OR = 3.34, 95% CI:1.48-7.54; P = 0.004) will significantly increase the occurrence of CRVT. However, there is no significant correlation between sex, congenital heart disease, bedridden state, sepsis, mechanical ventilation, anticoagulation therapy, insertion site (left), and CRVT. CONCLUSION: Our research results indicate that history of venous thrombosis, cancer, infection and multilumina are possible risk factors for CRVT, and corresponding preventive measures should be taken clinically.

[Research progress of refractory ceramic fiber's damage to human respiratory system].

Refractory ceramic fibers (RCFs) , as the main substitute for asbestos, are widely used because of their high temperature resistance and good thermal insulation. In the air of its production and use places, RCFs are inhalable fibers that are easy to deposit in the lungs. The results of a number of epidemiological studies and a variety of toxicological methods have shown that RCFs are related to the occurrence of lung diseases. This article reviews the four aspects of RCFs-induced pleural thickening, pulmonary fibrosis, lung function damage, tumor and genetic damage, and looks forward to the prospects of RCFs on respiratory system damage related research.

LncRNA DANCR promotes ATG7 expression to accelerate hepatocellular carcinoma cell proliferation and autophagy by sponging miR-222-3p.

OBJECTIVE: LncRNA differentiation antagonizing non-protein coding RNA (DANCR) is an oncogene in various malignant cancers, including hepatocellular carcinoma (HCC). Autophagy is an intracellular self-digestion mechanism that accelerates the progression of HCC via promoting cell survival. However, the role of lncRNA DANCR in HCC, and the mechanism of lncRNA DANCR in the regulation of autophagy in HCC remains unknown. Therefore, the aims of this study are the investigation of the role of lncRNA DANCR in HCC, and the exploration of the molecular mechanism of lncRNA DANCR in regulating autophagy of HCC cells. PATIENTS AND METHODS: In this study, the expression of lncRNA DANCR, miR-222-3p, and autophagy-related gene 7 (ATG7) was detected by qRT-PCR. The cell proliferation and colony formation were measured by Cell Counting Kit-8 (CCK-8) assay and colony formation assay. And the autophagic flux was evaluated by mRFP-GFP-LC3B reporter. The autophagy related proteins were analyzed by Western blotting. Besides, the relationship between lncRNA DANCR and miR-222-3p, as well as between miR-222-3p and ATG7, was determined by Dual-Luciferase reporter system. RESULTS: We found high expression of lncRNA DANCR and ATG7, and low expression of miR-222-3p in HCC tissues and cell lines. And lncRNA DANCR positively correlated with poor survival of HCC patients. Moreover, the knockdown of lncRNA DANCR inhibited cell proliferation and autophagy of HCC cells. And we predicted and proved that lncRNA DANCR induced cell proliferation, colony formation and autophagy by increasing ATG7 and suppressing miR-222-3p. CONCLUSIONS: Our study demonstrates the promoting role of lncRNA DANCR in HCC, and indicates the regulatory effects of lncRNA DANCR on regulating autophagy of HCC.

Heterogeneity of enhancement kinetics in dynamic contrast-enhanced MRI and implication of distant metastasis in invasive breast cancer.

AIM: To investigate the heterogeneity of enhancement kinetics for breast tumour in order to demonstrate the predictive power of dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) features for distant metastasis (DM) in invasive breast cancer. MATERIALS AND METHODS: Time-signal intensity curve (TIC) patterns from 128 patients with invasive breast cancer were analysed by a pixel-based DCE-MRI analysis. This MRI technique enabled pixels with varying TIC patterns (persistent, plateau, washout and non-enhancement) to be categorised semi-automatically and the percentage of different TIC patterns in each breast tumour to be calculated. The percentage of TIC patterns was compared between the DM and non-DM groups. DM-free survival was estimated using Kaplan-Meier survival analysis. RESULTS: This study demonstrated a larger percentage of persistent TIC and non-enhancement TIC was associated with DM in invasive breast cancer. The cut-off values of persistent TIC and non-enhancement TIC were 22.5% and 2.5%. Combining TIC patterns and traditional predictors (tumour size and axillary lymph node status) can improve the prediction efficiency. The multivariable model yielded an area under the receiver operating characteristic curve (AUC) of 0.87 with 0.70 sensitivity and 0.87 specificity in leave-one-out cross-validation (LOOCV). These predictors showed significant differences in DM-free survival by Kaplan-Meier analysis. CONCLUSION: This study shows that breast tumours with higher heterogeneity are more likely to metastasise, and pixel-based TIC analysis has utility in predicting distant metastasis of invasive breast cancer.

Urine-derived stem cells for the therapy of diabetic nephropathy mouse model.

OBJECTIVE: Diabetic nephropathy (DN) is one of the most representative diabetic microangiopathy complications. So far, there have been no satisfactory therapeutic strategies, and the injection of stem cells provides a target for DN therapy. PATIENTS AND METHODS: Urine-derived stem cells (USCs) were obtained from 9 healthy men. 24 mice were randomly and equally divided into control group, DN model group, DN+hUSC group (treated with USCs for 3 times). Hematoxylin-eosin (HE) and Masson staining were used to detect histological changes of kidney injury. Creatinine and blood urea nitrogen (BUN) were measured to assess renal function. Besides, myofibroblast accumulation, macrophage infiltration, cell proliferation, and oxidative stress were detected by immunohistochemical analysis. RESULTS: Compared with DN model group, DN+hUSC group showed lower function loss, cell infiltration, and oxidative stress, as well as less renal fibrosis, histological damage, and cell proliferation. CONCLUSIONS: USC can alleviate inflammation and oxidative stress, reduce renal interstitial fibrosis, improve renal tissue structure and protect renal function through paracrine effect.

[Cervial vestibular-evoked myogenic potential induced by galvanic vestibular stimulation in normal people].

Objective:To establish a new method for detecting vestibular function by testing cervical vestibular-evoked myogenic potential induced by galvanic vestibular stimulation in normal population. Method:Twenty normal ears were tested for cervical vestibular evoked myogenic potential induced by galvanic vestibular stimulation. SPSS 18.0 software was used to analyze the obtained data. Result:In all healthy subjects mastoid-forehead galvanic vestibular stimulation produced a positive-negative biphasic EMG responses on SCM ipsilateral to the cathodal electrode. The latency of p13 was（11.52±3.05） ms. The latency of n23 was（15.31±3.38） ms. The amplitude of p13-n23 was（40.55±27.93） µV. The interval of p13-n23 was（3.53±1.38） ms. The interaural asymmetry ratio（AR, %） of p13, n23 latency, the amplitude and interval were respectively（6.96±6.79）%, （6.47±5.93）%, （28.08±26.42）% and （16.61±11.11）%. There was no significant difference in all parameters between the right and left ears of all subjects. Conclusion:The value of cervical vestibular-evoked myogenic potential induced by galvanic vestibular stimulation in normal subjects can be established to explore methods for diagnosis, treatment and researching mechanism of auditory neuropathy and vestibular neuropathy.

[Health literacy and cancer prevention].

This article reviews the literature on health literacy and cancer prevention, screening, treatment and prognosis. A large number of studies have shown that health literacy is positively correlated with public cancer risk awareness, cancer knowledge awareness rate, and cancer screening behavior, and positively correlated with the health service resource utilization ability of cancer patients, treatment compliance, and quality of life, and negatively correlated with the unhealthy lifestyle such as smoking and drinking. Some studies have problems such as small sample size, limited population, and inappropriate design. Some studies do not support the conclusions above. Therefore, multi-center, large-scale clinical studies and cohort studies should be conducted to provide more robust evidence for the relationship between the health literacy and cancer prevention, screening, and treatment.

[Association between low handgrip strength and air pollution among people aged 50 years and over].

Objective: To examine the association between long-term exposure to ambient PM(2.5) combined with indoor air pollution and handgrip strength among people aged 50 and over. Methods: Data were from the first wave of World Health Organization Study on global AGEing and adult health in China. Ambient annual concentration of PM(2.5) was estimated by using the satellite data we also investigated the use of fuels and chimneys as indoor air pollution. A two-level (individual level and community level) linear model was applied to examine the association between long-term exposure to ambient PM(2.5) combined with indoor air pollution and the handgrip strength. Results: A total of 13 175 individuals aged 50 years and over were included for analysis. The handgrip strength was (26.67±0.54) kg. Ambient PM(2.5) was found to be significantly associated with the risk of decreased handgrip strength. Outdoor PM(2.5) concentration was negatively correlated with handgrip strength (ß=-0.23, 95%CI: -0.31 - -0.14) decrease in handgrip strength after adjusting for gender, age, residence, education, household assets, intake of vegetables and fruits, smoking and drinking, physical activity. In rural area, compared to those who used solid fuel, use of clean fuel increased (ß=1.41, 95%CI: 0.36-2.46) handgrip strength. But in urban area, we did not find any statistically significant association between the use of clean fuel and handgrip strength (ß=0.19, 95%CI: -0.95-1.32). Conclusion: This study found that long-term exposure to ambient PM(2.5) combined with indoor air pollution was significantly associated with low handgrip strength among people aged 50 years and over, this suggested that ambient PM(2.5) might serve as one of the risk factors for low physical function seen in the people aged 50 years and over.

[Correlation between breast density and age, breast cancer molecular subtype].

Objective: To explore the correlation between breast density and breast cancer molecular subtype. Methods: The data of 1 407 cases of breast cancer immunohistochemistry and preoperative mammography in Tianjin Medical University Cancer Institute and Hospital from August 2015 to August 2016 were retrospectively analyzed. All cases were divided into four groups: <45 years old, 45-54 years old, 55-64 years old and ≥ 65 years old, and all groups were divided into four molecular subtypes: Luminal A, Luminal B, HER2-enriched and triple-negative according to immunohistochemical results. Person correlation test was used to analyze the correlation between breast density and age; Kruskal-Wallis test was used to analyze the difference in breast density between different molecular subtypes. Results: There was a negative correlation between breast density and age (r=-0.55, P<0.01). Descriptive analysis results showed that the mean breast density of<45 years old, 45-54 years old, 55-64 years old and ≥65 years old was 20.0%±7.3%, 16.2%±8.4%, 10.5%±5.2%, and 7.9%±3.2%, respectively. The mean breast density of Luminal A, Luminal B, HER2-enriched and triple-negative was 14.0%±8.1%, 14.5%±7.6%, 14.8%±7.7% and 13.2%±7.3%, respectively. Kruskal-Wallis test showed significant statistical differences in breast density among molecular subtypes in the group of<45 years old (P<0.05), while no statistical significance in other groups (all P>0.05). Conclusion: There is correlation between breast density and breast cancer molecular subtype in young patients, but whether breast density has a potential evaluation effect on breast cancer molecular subtype still needs to be further studied.

[The experimental study on endoplasmic reticulum stress-participated outer hair cell apoptosis in cadherin 23 gene mutant mice].

Objective: To test the mechanism and upstream pathway of outer hair cell apoptosis in Cadherin 23 (Cdh23) gene mutant mice. Method: The mutant Cdh23(erl/erl)(erl) mice were collected as the study group, while the C57BL/6J (B6) mice were chosen as the control group. A total of 70 mice per group were used in this study. The study group and control group underwent auditory-evoked brainstem response (ABR) tests at the same age. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect outer hair cell(OHC) apoptosis. The qRT-PCR was conducted to test the expression of ER stress markers immunoglobulin-binding protein (BiP) and C/EBP homologous protein (CHOP) mRNA. The expression and location of BiP and CHOP protein in OHC were detected by immunostaining. The expression of BiP protein in cochleae was identified by Western blot. The expression and location of CDH23 protein in OHC were discovered by immunostaining. Results: The ABR thresholds in erl mice were significantly higher than those in B6 mice at the age of 1 and 3 months (both P<0.05). The surface preparation with TUNEL staining confirmed OHC apoptosis in erl mouse cochleae which showed a higher TUNEL positive cell ratio than B6 mouse(t=11.291, P<0.01). The ER stress marker Bip and Chop mRNA were upregulated in the erl mouse inner ear, when compared with those in the B6 mouse(both P<0.05). The BiP protein extracted from the erl mouse cochleae was significantly higher than that of B6 mouse measured by Western blot (t=3.66, P=0.02). Immunostaining showed that BiP and CHOP were highly detected in the OHC in erl mouse cochleae, and was mainly detected in the perinuclear region of OHC. However, a bare BiP and CHOP signal were shown in B6 mouse cochleae. The CDH23 protein was specifically localized at the top of the OHC in B6 mice, indicating the localization of the tip links in hair bundle stereocilia. On the contrary, the CDH23(erl) protein was found to be localized from the top to the nuclei of the OHC in erl mice. Portions of the CDH23(erl) proteins failed to reach the top of the hair bundles and remained in the OHC cytoplasm. Conclusion: As the downstream response of the Cdh23 gene mutation, portions of the mutant CDH23(erl) protein was accumulated in ER lumen resulting in the increase of ER loading and ultimately triggered ER stress and hair cell apoptosis in erl mouse cochleae.

[Advance in research of delayed endolymphatic hydrops].

Delayed endolymphatic hydrops (DEH) is a disease entity first described by Kamei and named by Schuknecht, defined as profound sensorineural deafness at early stage and after several years started to appear clinical feature of endolymphatic hydrops such as vertigo, aural fullness like Meniere's disease or fluctuating hearing loss in the contralateral ear . DEH can be classified into ipsilateral type, contralateral type and bilateral type. Although DEH has low incident rate, there are many kinds of etiology and audiology and vestibular tests. Up to now, a lot of literatures about etiology, diagnose, clinical manifestation, relevant examination and therapy of DEH were published abroad. In this review, we will make a summary of research status of DEH.

Genetic characterization of cysteine-rich type-b avenin-like protein coding genes in common wheat.

The wheat avenin-like proteins (ALP) are considered atypical gluten constituents and have shown positive effects on dough properties revealed using a transgenic approach. However, to date the genetic architecture of ALP genes is unclear, making it impossible to be utilized in wheat breeding. In the current study, three genes of type-b ALPs were identified and mapped to chromosomes 7AS, 4AL and 7DS. The coding gene sequence of both TaALP-7A and TaALP-7D was 855 bp long, encoding two identical homologous 284 amino acid long proteins. TaALP-4A was 858 bp long, encoding a 285 amino acid protein variant. Three alleles were identified for TaALP-7A and four for TaALP-4A. TaALP-7A alleles were of two types: type-1, which includes TaALP-7A1 andTaALP-7A2, encodes mature proteins, while type-2, represented byTaALP-7A3, contains a stop codon in the coding region and thus does not encode a mature protein. Dough quality testing of 102 wheat cultivars established a highly significant association of the type-1 TaALP-7A allele with better wheat processing quality. This allelic effects were confirmed among a range of commercial wheat cultivars. Our research makes the ALP be the first of such genetic variation source that can be readily utilized in wheat breeding.

[Molecular mechanisms of resistance to phosphatidyl inositol 3-kinase inhibitors in triple-negative breast cancer cells].

OBJECTIVE: To explore the molecular mechanisms of resistance to phosphatidyl inositol 3-kinase (PI3K) inhibitors in triple-negative breast cancer (TNBC) cells. METHODS: HCC70 cells (TNBC) were transfected with siFZD7, siWANT5B or siGSK3 using lipofectamine 2000 transfection reagent. The expression levels of key proteins of WNT/ß-catenin and PI3K/AKT/mTOR pathways were determined by Western blot analysis. After HCC70, MCF-7 (ER-positive) and SK-BR3 (HER2-positive) cells were treated with PI3K/AKT/mTOR inhibitors, the inhibition rates of cell proliferation were measured by MTT assay, and half maximal inhibitory concentrations (IC50) were calculated. The altered activities of WNT/ß-catenin and PI3K/AKT/mTOR proteins were detected by Western blot and luciferase report gene assay, respectively. The nuclear translocation of ß-catenin protein was examined by immunofluorescence assay. Xenograft nude mouse model was used to evaluate the tumorigenicity of breast cancer cells treated with BKM120 in vivo. The expression levels of p-LRP6, p-4EBP1 and ß-catenin proteins in the tumor tissues were determined by immunohistochemical staining. RESULTS: The expression levels of FZD7, WANT5B and GSK3 proteins were significantly reduced in the HCC70 cells transfected with the target siRNAs. Meanwhile, the activity of WNT/ß-catenin was enhanced and PI3K/AKT/mTOR pathway was inhibited. PI3K/AKT/mTOR inhibitors suppressed MCF-7 and SK-BR3 cell proliferation. The IC50 of GDC-094, BKM120, XL147, perifosine, everolimus, and BEZ235 in MCF-7 cells were 0.46 mmol/L, 1.44 mmol/L, 4.34 mmol/L, 11.35 µmol/L, 53.71 µmol/L and 12.87 µmol/L respectively, and 0.63 mmol/L, 0.58 mmol/L, 3.74 mmol/L, 13.22 µmol/L, 60.00 µmol/L and 11.38 µmol/L in the SK-BR3 cells, respectively. The results of luciferase report gene assay showed that the luciferase activities in HCC70, MCF-7 and SK-BR3 cells treated with BKM120 were 1.75±0.05, 1.13±0.02 and 0.43±0.01, respectively. The luciferase activities in HCC70 and SK-BR3 cells were significantly different from that of the control cells (1.00±0.02, P<0.05). The immunohistochemical analysis showed that BKM120 inhibited mTOR activity, and the enhanced WNT/ß-catenin activity reversed the phenotype of inhibitory mTOR induced by BKM120. BKM120 suppressed the tumorigenic ability of MCF-7 and SK-BR3 cells in vivo, but had no effect on cultured HCC70 cells. The immunohistochemical analysis showed nuclear translocation of ß-catenin protein and increased expression level of p-LRP-6 protein in transplanted tumor tissues from HCC70 cells treated with BKM120, increased the level of p-LRP-6 protein, and no changes of p-4EBP1 protein expression. However, no nuclear translocation of ß-catenin protein and no decrease of p-LRP6 and p-4EBP1 protein levels in the transplanted tumor tissue of MCF-7 cells after treatment with BKM120. CONCLUSIONS: The triple-negative breast cancer HCC70 cells have drugs-resistance to PI3K inhibitors. The WNT/ß-catenin signaling pathway may regulate the PI3K/AKT/mTOR pathway, therefore, inducing the drug-resistance of TNBC cells to PI3K inhibitors.

IL-9(+) IL-10(+) T cells link immediate allergic response to late phase reaction.

The mechanism underlying late-phase allergic reactions (LPR) remains incompletely understood. This study aimed to investigate the role of a newly described subset of T cells, interleukin (IL)-9(+) IL-10(+) T cells, in the pathogenesis of LPR. Using a T helper type 2 (Th2) inflammatory mouse model, we examined the frequency of IL-9(+) IL-10(+) T cells in the jejunum by immunohistochemistry. The LPR in the jejunum was observed afterwards. The cytokine profile of IL-9(+) IL-10(+) T cells was characterized and the major cytokine that plays the critical role in the initiation of LPR was investigated. Abundant IL-9(+) IL-10(+) T cells as well as inflammatory cell extravasation in the jejunal sections were observed in sensitized mice 48 h after specific antigen challenge. IL-9(+) IL-10(+) T cells expressed high levels of macrophage inflammatory protein 1 (MIP1) that could be enhanced by T cell receptor activation. MIP1 facilitated macrophage extravasation in local tissue. Macrophage-derived MIP2 contributed to neutrophil infiltration in the intestine in LPR. Pretreatment with anti-MIP antibody inhibited the LPR in the intestine. IL-9(+) IL-10(+) T cells play an important role in LPR. This subset of T cells has the potential to be a novel therapeutic target in the treatment of LPR and LPR-related inflammation.

Mitochondrial function is altered in articular chondrocytes of an endemic osteoarthritis, Kashin-Beck disease.

OBJECTIVE: Kashin-Beck disease (KBD) is an endemic degenerative osteoarthritis (OA) associated with extracellular matrix degradation and chondrocyte necrosis in the articular and growth plate cartilage. The role of mitochondria in degenerative diseases is widely recognized but its function in KBD is unknown. The aim of this investigation was to evaluate mitochondrial function to understand the mitochondria-mediated caspase activation and apoptosis in adult KBD chondrocytes. METHODS: Mitochondrial function was evaluated by analyzing the activities of respiratory chain enzyme complexes and citrate synthase (CS), intracellular adenosine triphosphate (ATP) contents, as well as changes in mitochondrial membrane potential (DeltaPsim). Apoptotic cell death was evaluated by analyzing the cytochrome c release from mitochondria to the cytosol, caspase-9 and 3 activities, and the apoptosis rate of KBD articular chondrocytes. RESULTS: Activities of complexes II, III, IV and V were reduced in KBD articular chondrocytes compared with cells from normal controls. However, the mitochondrial mass was increased in KBD samples. Cultured KBD chondrocytes had a reduction of cellular ATP levels and contained a higher proportion of cells with de-energized mitochondria. Mitochondrial cytochrome c release and activation of caspase-9 and 3 were also observed. The percentages of positive apoptotic chondrocytes from the KBD patient group stained by Hoechst nuclear stain and Annexin V/PI for flow cytometry exhibited higher levels than that of the healthy controls. CONCLUSION: These findings suggest the involvement of mitochondrial function and apoptotic cell death in the pathophysiology of KBD. The dysfunction of the mitochondria may play an important role in KBD articular chondrocytes apoptosis.

Intracellular calcium elevation induced by extracellular application of cyclic-ADP-ribose or oxytocin is temperature-sensitive in rodent NG108-15 neuronal cells with or without exogenous expression of human oxytocin receptors.

ADP-ribosyl cyclase and/or CD38 are activated after oxytocin receptor stimulation in the hypothalamus and pituitary in adult mice, leading to facilitation of oxytocin secretion. Although cyclic adenosine 5'-diphosphoribose (cADPR) primarily acts as an intracellular second messenger, it has been suggested that extracellular cADPR stimulates intracellular ryanodine receptors after internalisation via the nucleotide-transporting capacity of CD38 in fibroblasts and astrocytes. However, little is known about whether extracellular cADPR activates neurones. To address this question, we used a model neuronal cell line, NG108-15 mouse neuroblastoma x rat glioma hybrid cells possessing CD38 but not oxytocin receptors, and measured cytosolic free calcium concentrations ([Ca(2+)](i)). Extracellular application of cADPR to NG108-15 cells elevated [Ca(2+)](i) at 35 degrees C. The elevation was significantly enhanced when measured at 40 degrees C. The cADPR and heat-induced [Ca(2+)](i) increase were blocked under extracellular Ca(2+)-free conditions and by 2-aminoethoxydiphenyl borate, an antagonist of melastatin-related transient receptor potential channel 2 (TRPM2) cation channels. Reverse transcriptation-polymerase chain reaction analyses indicated that TRPM2 channels were expressed in NG108-15 cells. Application of oxytocin elevated [Ca(2+)](i) in NG108-15 cells transformed to transiently express cloned human oxytocin receptors. The oxytocin-induced [Ca(2+)](i) response was also enhanced by heat. These results indicate that the extracellular application of cADPR, together with heat, activates cation influx downstream of oxytocin receptor signalling in NG108-15 neuronal cells, and suggest the possible involvement of TRPM2 channels in oxytocin release in the mammalian brain.