Establishment of a prognostic risk prediction modelfor non-small cell lung cancer patients with brainmetastases: a retrospective study.

Background: Patients with non-small cell lung cancer (NSCLC) who develop brain metastases (BM) have a poor prognosis. This study aimed to construct a clinical prediction model to determine the overall survival (OS) of NSCLC patients with BM. Methods: A total of 300 NSCLC patients with BM at the Yunnan Cancer Centre were retrospectively analysed. The prediction model was constructed using the least absolute shrinkage and selection operator-Cox regression. The bootstrap sampling method was employed for internal validation. The performance of our prediction model was compared using recursive partitioning analysis (RPA), graded prognostic assessment (GPA), the update of the graded prognostic assessment for lung cancer using molecular markers (Lung-molGPA), the basic score for BM (BSBM), and tumour-lymph node-metastasis (TNM) staging. Results: The prediction models comprising 15 predictors were constructed. The area under the curve (AUC) values for the 1-year, 3-year, and 5-year time-dependent receiver operating characteristic (curves) were 0.746 (0.678-0.814), 0.819 (0.761-0.877), and 0.865 (0.774-0.957), respectively. The bootstrap-corrected AUC values and Brier scores for the prediction model were 0.811 (0.638-0.950) and 0.123 (0.066-0.188), respectively. The time-dependent C-index indicated that our model exhibited significantly greater discrimination compared with RPA, GPA, Lung-molGPA, BSBM, and TNM staging. Similarly, the decision curve analysis demonstrated that our model displayed the widest range of thresholds and yielded the highest net benefit. Furthermore, the net reclassification improvement and integrated discrimination improvement analyses confirmed the enhanced predictive power of our prediction model. Finally, the risk subgroups identified by our prognostic model exhibited superior differentiation of patients' OS. Conclusion: The clinical prediction model constructed by us shows promise in predicting OS for NSCLC patients with BM. Its predictability is superior compared with RPA, GPA, Lung-molGPA, BSBM, and TNM staging.

Identification of epigenetic modifications mediating the antagonistic effect of selenium against cadmium-induced breast carcinogenesis.

The antagonistic effect of selenium (Se) against cadmium (Cd)-induced breast carcinogenesis was reported, but underlying mechanisms were unclear. The aim of this study was to identify the epigenetically regulated genes and biological pathways mediating the antagonistic effect. We exposed MCF-7 cells to Cd and Se alone or simultaneously. Cell proliferation was assessed by MTT assay, and differential epigenome (DNA methylation, microRNA, and long non-coding RNA) was obtained by microarrays. We cross-verified the epigenetic markers with differential transcriptome, and the ones modulated by Cd and Se in opposite directions were regarded to mediate the antagonistic effect. The epigenetically regulated genes were validated by using gene expression data in human breast tissues. We further assessed the biological functions of these validated genes. Our results showed that Se alleviated the proliferative effect of Cd on MCF-7 cell. A total of 10 epigenetically regulated genes were regarded to mediate the antagonistic effect, including APBA2, KIAA0895, DHX35, CPEB3, SVIL, MYLK, ZFYVE28, ABLIM2, GRB10, and PCDH9. Biological function analyses suggested that these epigenetically regulated genes were involved in multiple cancer-related pathways, such as focal adhesion and PI3K/Akt pathway. In conclusion, we provided evidence that Se antagonized the Cd-induced breast carcinogenesis via epigenetic modification and revealed the critical pathways.

Differential epigenetic profiles induced by sodium selenite in breast cancer cells.

OBJECTIVES: Selenium (Se) was a potential anticancer micronutrient with proposed epigenetic effect. However, the Se-induced epigenome in breast cancer cells was yet to be studied. METHODS: The profiles of DNA methylation, microRNA (miRNA), long non-coding RNA (lncRNA), and message RNA (mRNA) in breast cancer cells treated with sodium selenite were examined by microarrays. We verified the epigenetic modifications by integrating their predicted target genes and differentially expressed mRNAs. The epigenetically regulated genes were further validated in a breast cancer cohort by associating with tumor progression. We conducted a series of bioinformatics analyses to assess the biological function of these validated genes and identified the critical genes. RESULTS: The Se-induced epigenome regulated the expression of 959 genes, and 349 of them were further validated in the breast cancer cohort. Biological function analyses suggested that these validated genes were enriched in several cancer-related pathways, such as PI3K/Akt and metabolic pathways. Based on the degrees of expression change, hazard ratio difference, and connectivity, NEDD4L and FMO5 were identified as the critical genes. CONCLUSIONS: These results confirmed the epigenetic effects of sodium selenite and revealed the epigenetic profiles in breast cancer cells, which would help understand the mechanisms of Se against breast cancer.

Differential epigenetic and transcriptional profile in MCF-7 breast cancer cells exposed to cadmium.

Cadmium (Cd) has been confirmed to be associated with breast carcinogenesis, but the mechanism was not clarified yet. Given that epigenetic modification was speculated as underlying mechanism, we examined the differential epigenome caused by Cd in breast cancer cells. Profiles of DNA methylation, microRNA (miRNA), long non-coding RNA (lncRNA), and message RNA (mRNA) were derived from Cd-treated and untreated MCF-7 breast cancer cells by microarray. We identified 997 target genes epigenetically regulated by Cd through cross-verification with the differential epigenome and transcriptome, and 400 of them were further validated in a breast cancer cohort. Biological function analyses suggested that several pathways were involved in Cd-induced breast carcinogenesis, such as Wnt signaling, metabolism, and human papilloma virus (HPV) infection. TXNRD1 and CCT3 were further identified as the critical genes based on the degree of expression change, hazard ratio difference, and connectivity. The present study revealed that Cd epigenetically regulated several pathways involving in breast carcinogenesis, particularly the Wnt signaling and metabolic pathways, among which TXNRD1 and CCT3 might play critical roles. It was also suggested that Cd and HPV infection might jointly participate in breast tumorigenesis.

Anxiety and depression in patients with gastroesophageal reflux disease and their effect on quality of life.

AIM: To explore the role of psychological factors in gastroesophageal reflux disease (GERD) and their effect on quality of life (QoL) of GERD patients. METHODS: A total of 279 consecutive patients with typical symptoms and 100 healthy controls were enrolled in the study. All of the participants were evaluated with the Zung Self-Rating Anxiety Scale (ZSAS), the Zung Self-Rating Depression Scale (ZSDS) and the SF-36 questionnaire. The scores for anxiety, depression and QoL of the two groups were analyzed. The correlation between psychological factors and QoL was also analyzed. RESULTS: Compared with healthy controls (34.70 ± 8.00), the scores of ZSAS in the non-erosive reflux disease (NERD) group (48.27 ± 10.34) and the reflux esophagitis (RE) group (45.38 ± 10.27) were significantly higher (P < 0.001). The mean ZSAS score of the NERD group was significantly higher than that of the RE group (P = 0.01). Compared with healthy controls (37.61 ± 8.44), the mean ZSDS scores were significantly higher in the NERD group (49.65 ± 11.09, P < 0.001) and the RE group (46.76 ± 11.83, P < 0.001). All dimensions of the SF-36 form were negatively correlated with the SAS and SDS scores in patients with NERD and RE (P < 0.05). According to the SF-36 form, vitality, mental health and social functioning were significantly correlated with symptoms of depression in patients with NERD and RE. General health was obviously affected by symptoms of depression in patients with NERD (P < 0.05). CONCLUSION: Anxiety and depression may play an important role in the occurrence of GERD and especially that of NERD. The QoL of patients with GERD is reduced by anxiety and depression.

Magnetic solid-phase extraction combined with graphite furnace atomic absorption spectrometry for speciation of Cr(III) and Cr(VI) in environmental waters.

A new approach of magnetic solid phase extraction (MSPE) coupled with graphite furnace atomic absorption spectrometry (GFAAS) has been developed for the speciation of Cr(III) and Cr(VI) using zincon-immobilized silica-coated magnetic Fe3O4 nanoparticles (Zincon-Si-MNPs) as the MSPE absorbent. Cr(III) was quantitatively reserved on the absorbent at pH 9.1 while total Cr was reserved at pH 6.5. The absorbed Cr species were eluted by using 2 mol/L HCl and detected by GFAAS. The concentration of Cr(VI) could be calculated by subtracting Cr(III) from total Cr. All the parameters affecting the separation and extraction efficiency of Cr species such as pH, extraction time, concentration and volume of eluent, sample volume and influence of co-existing ions were systematically examined and the optimized conditions were established accordingly. The detection limit (LOD) of the method was 0.016 and 0.011 ng mL(-1) for Cr(III) and Cr(VI), respectively, with the enrichment factor of 100 and 150. The precisions of this method (Relative standard deviation, RSD, n=7) for Cr(III) and Cr(VI) at 0.1 ng mL(-1) were 6.0% and 6.2%, respectively. In order to validate the proposed method, a certified reference material of environmental water was analyzed, and the result of Cr speciation was in good agreement with the certified value. This MSPE-GFAAS method has been successfully applied for the speciation of Cr(III) and Cr(VI) in lake and tap waters with the recoveries of 88-109% for the spiked samples. Moreover, the MSPE separation mechanism of Cr(III) and Cr(VI) based on their adsorption-desorption on Zincon-Si-MNPs has been explained through various spectroscopic characterization.

[Effects of low-dose insulin on oxidation-reduction in rat organs after major burns].

OBJECTIVE: To explore the effects of low-dose insulin on oxidation-reduction of heart and kidney in rats immediately after severe burns. METHODS: Twenty four male Sprague-Dawley (SD) rats were randomly divided into 3 groups of sham burn, burn and treatment (n = 8 each). The sham burn group was placed into 37 °C warm water for 15 seconds to simulate burn process and received no fluid replacement. The burn and treatment groups were immersed into (95 ± 0.5) °C hot water for 15 seconds to make a rat model of 30% total burn surface area, III degree burn injury and immediately received an intraperitoneal injection of physiological saline (40 ml/kg). At the same time, a subcutaneous injection of insulin (1.0 U×kg(-1)×d(-1)) was given in the treatment group and a subcutaneous injection of same-volume physiological saline in the burn group. The rats were sacrificed after 24 post-scald hours (PSH). Abdominal aortic blood was collected for an analysis of blood glucose. The oxidation and antioxidation parameters of heart and kidney, such as malondialdehyde (MDA), xanthine oxidase (XO), myeloperoxidase (MPO), total superoxide dismutase (T-SOD), superoxide dismutase 1, 2 (SOD1, 2), catalase (CAT) and glutathione peroxidase (GPx), were detected by spectrophotometry. RESULTS: Compared with the sham burn group, MDA content, XO and MPO activities of heart and kidney were significantly higher in the burn group (all P < 0.05); in the treatment group, MDA content and XO activity of heart were significantly lower than the burn group ((0.85 ± 0.07) vs (1.11 ± 0.07) nmol/mg, (69.72 ± 1.94) vs (77.21 ± 2.10) U/g) while the MPO activities of heart and kidney were significantly lower (all P < 0.05).compared with the sham burn group, the activities of T-SOD, CAT, GPx of heart and kidney were significantly lower in the burn group, SOD1 activity of kidney was significantly lower, but SOD2 activity of kidney was significantly higher while SOD2 activity of heart was significantly lower (all P < 0.05); Compared with the burn group, the activities of T-SOD and SOD1 of heart in the treatment group were significantly higher ((83.5 ± 2.5) vs (79.6 ± 3.2), (62.8 ± 2.3) vs (58.8 ± 3.0) U/mg), CAT and GPx activity of heart and kidney were significantly higher (all P < 0.05). Compared with the sham burn group, blood glucose in the burn and treatment group were significantly higher ((7.81 ± 0.30), (7.19 ± 0.22) vs (6.30 ± 0.24) mmol/L) and blood glucose in the treatment group was significantly lower than the burn group((7.81 ± 0.30) mmol/L)(all P < 0.05). CONCLUSIONS: During an early stage, a low-dose insulin may intervene in heart tissue lipid peroxidation of severely burned rats. And differences exist in the effects of oxidation-reduction between heart and kidney.

Zincon-immobilized silica-coated magnetic Fe3O4 nanoparticles for solid-phase extraction and determination of trace lead in natural and drinking waters by graphite furnace atomic absorption spectrometry.

A new protocol using zincon-immobilized silica-coated magnetic Fe(3)O(4) nanoparticles (Zincon-Si-MNPs) as solid-phase extraction (SPE) medium has been developed for the separation and preconcentration of trace lead in water. Various parameters such as pH, extraction time, concentration and volume of eluent, sample volume, and influence of co-existing ions have been investigated in order to establish the optimum conditions for the determination of lead in combination with graphite furnace atomic absorption spectrometry (GFAAS). The detection limit (LOD) of the proposed method for lead based on an enrichment factor of 200 was 10 ng L(-1). The relative standard deviations (RSDs, n=5) were 8.3%, 7.8% and 9.2%, respectively, at 5, 0.5 and 0.05 ng mL(-1) levels. This method has been successfully applied to the analysis of trace lead in natural and drinking water samples and the recoveries for the spiked samples were in the range of 84-104%.

[Reversed changes of the 15-lipoxygenase product lipoxin A4 and the 5-lipoxygenase product leukotriene C4 in children with asthma].

OBJECTIVE: To investigate the expressions of 15- and 5-lipoxygenases in leukocytes and the changes of the levels of blood lipoxin A4 (LXA4) and leukotriene C4 (LTC4) in children with asthma. METHODS: The mRNA levels of 15- and 5-lipoxygenases in leukocytes were assessed by RT-PCR, and the levels of blood LXA4 and LTC4 were determined by ELISA, in 106 children with mild, moderate and severe asthma. Forty healthy children served as the controls. RESULTS: In children with mild, moderate and severe asthma, the relative mRNA levels of 15-lipoxygenase in leukocytes were 1.78 ± 0.56, 1.28 ± 0.45 and 0.58 ± 0.22 (F = 16.72, P < 0.01), respectively, and all were higher than that of the controls (0.26 ± 0.12, P < 0.05). The levels of blood LXA4 were (5.52 ± 1.97), (1.86 ± 0.72) and (0.81 ± 0.36) µg/L (F = 22.59, P < 0.01), respectively, decreasing with the severity of asthma, and all were higher than that of the controls [(0.04 ± 0.01) µg/L, P < 0.05]. There was a positive correlation between PEF, FEV(1) and blood LXA4. The relative levels of 5-lipoxygenase mRNA in leukocytes were 0.26 ± 0.12, 0.79 ± 0.34 and 1.21 ± 0.52, respectively in children with asthma of mild, moderate and severe degree (F = 18.64, P < 0.01), which showed an increase with the severity of the disease, and all of which were higher than that of the controls (0.12 ± 0.05, P < 0.05). The levels of blood LTC4 were (22.4 ± 8.2), (54.6 ± 28.4) and (118.7 ± 41.1) ng/L (F = 25.91, P < 0.01), respectively, also showing an increase with the severity of asthma, and were higher than that of the controls [(6.8 ± 2.5) ng/L, P < 0.05]. There was a negative correlation between PEF, FEV1 and blood LTC4. CONCLUSION: The reversed changes of 15-lipoxygenase product LXA4 and 5-lipoxygenase product LTC4 in children with asthma of mild, moderate and severe degree suggests that insufficiency of LXA4, an physiological antagonist to leukotrienes, and an overproduction of LTC4, may be involved in the pathogenesis of worsening of asthma in children.

[HLA-DM polymorphisms in Guizhou Han individuals].

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine genetic polymorphisms at HLA-DMA/DMB loci in 125 healthy unrelated Han individuals from Guizhou Province. We found that the frequencies of DMA*0101, DMA*0102, and DMA*0103 alleles were 0.720, 0.242, and 0.036, respectively. The frequencies of DMB*0101, DMB*0102, DMB*0103, and DMB*0104 alleles were 0.620, 0.156, 0.188, and 0.036, respectively. Common genotypes for DMA were DMA*0101/0101 and 0101/0102. Common genotypes for DMB were DMB*0101/0101, 0101/0102, and 0101/0103. These results showed that the genetic polymorphism of HLA-DM gene in Han individuals from Guizhou Province is distinct from other regions.

[Study on extraction technology of Mosla chinensis volatile oil].

OBJECTIVE: To study the techniques of organic solvent extraction method and supercritical CO2 extraction for extracting Mosla chinensis Maxim volatile oil (MCMVO), and to analyze its chemical compositions. METHODS: Organic solvent extraction method and supercritical CO2 extraction were used to extract the oil and GC-MS was used to analyze its chemical compositions. RESULTS: The best extraction techniques of organic solvent method to extract volatile oil should be divided into two steps, total time was 210 minutes, the total proportions of input of the solvent and raw materials was 8:1 (ml:g), and temprature was 65 degrees C, its extraction rate was 1.83%. The MCMVO was extracted by supercritical CO2, and its extraction rate was 3.4%. GC-MS was used to analyze volatile oil, the results suggested MCMVO contain thymol, carvacrol, P-cycmene, humulene etc. The thymol had the highest content accounts for 56.25%, and the carvacrol was 19.21%. The content of thymol and carvacrol of MCMVO extracted by organic solvent was 58.33% and 22.54%. CONCLUSION: the content of thymol and carvacrol extracted by organic solvent extraction method is higher than that extracted by supercritical CO2 extraction method. Chemical compositions extracted by organic solvent extraction method are less than extracted by supercritical CO2 extraction method. 10 compositions are firstly extracted by supercritical CO2 extraction method.

[Influence of anoxia/reoxygenation on immunofunction of endothelial cells and effect of intervention with yisheng injection on it].

OBJECTIVE: To explore the influence of ischemia/reperfusion (anoxia/reoxygenation) on immunofunction of endothelial cells (ECs) and effect of intervention with Yisheng injection (YSI, a pure natural medicine) on it. METHODS: Model of ECs induced by anoxia/reoxygenation was established to mimic ECs ischemia/reperfusion injury in vivo with human umbilical vein endothelial cell line ECV304. Then YSI was used to intervene the anoxia/reoxygenation process. Nuclear transcriptional factor-kappa B (NF-kappa B) was exhibited by fluorescent staining, HLA-ABC, HLA-DR, CD86 and CD54 were detected by flow cytometry. Mixed endothelial cell-lymphocyte reaction (MELR) was conducted to examine the proliferation of lymphocyte, production of IL-2 and percentage of apoptotic lymphocyte. RESULTS: Anoxia/reoxygenation made the ECV304 cell became round, shrunk and abscised, with increased plasma NF-kappa B, and changed from positive cytoplasm to positive nucleus. HLA-ABC, HLA-DR and CD86 on surface of cells increased but CD54 showed unchanged. MELR showed the incorporation of 3H-TdR and production of IL-2 increased significantly and the percentage of apoptotic lymphocyte decreased. After YSI intervention, the ECV304 cell shaped recovered, NF-kappa B expression didn't down-regulated, but the percentage of positive cells decreased, changed to positive dominant. Besides, reversal changes were shown in other parameters. CONCLUSION: Anoxia/reoxygenation influences some important immune related molecules in ECV304 cells, YSI could antagonizing these influences to maintain the immune function of endothelial cells in a relative normal manner.