L-carnitine reduces acute lung injury via mitochondria modulation and inflammation control in pulmonary macrophages.

Acute lung injury (ALI) or acute respiratory distress syndrome (ARDS) is a critical respiratory syndrome with limited effective interventions. Lung macrophages play a critical role in the pathogenesis of abnormal inflammatory response in the syndrome. Recently, impaired fatty acid oxidation (FAO), one of the key lipid metabolic signalings, was found to participate in the onset and development of various lung diseases, including ALI/ARDS. Lipid/fatty acid contents within mouse lungs were quantified using the Oil Red O staining. The protective effect of FAO activator L-carnitine (Lca, 50, 500, or 5 mg/mL) was evaluated by cell counting kit 8 (CCK-8) assay, real-time quantitative PCR (qPCR), ELISA, immunoblotting, fluorescence imaging, and fluorescence plate reader detection in lipopolysaccharide (LPS) (100 ng/mL)-stimulated THP-1-derived macrophages. The in vivo efficacy of Lca (300 mg/kg) was determined in a 10 mg/kg LPS-induced ALI mouse model. We found for the first time that lipid accumulation in pulmonary macrophages was significantly increased in a classical ALI murine model, which indicated disrupted FAO induced by LPS. Lca showed potent anti-inflammatory and antioxidative effects on THP-1 derived macrophages upon LPS stimulation. Mechanistically, Lca was able to maintain FAO, mitochondrial activity, and ameliorate mitochondrial dynamics. In the LPS-induced ALI mouse model, we further discovered that Lca inhibited neutrophilic inflammation and decreased diffuse damage, which might be due to the preservation of mitochondrial homeostasis. These results broadened our understanding of ALI/ARDS pathogenesis and provided a promising drug candidate for this syndrome.

L-carnitine reduces acute lung injury via mitochondria modulation and inflammation control in pulmonary macrophages

Acute lung injury (ALI) or acute respiratory distress syndrome (ARDS) is a critical respiratory syndrome with limited effective interventions. Lung macrophages play a critical role in the pathogenesis of abnormal inflammatory response in the syndrome. Recently, impaired fatty acid oxidation (FAO), one of the key lipid metabolic signalings, was found to participate in the onset and development of various lung diseases, including ALI/ARDS. Lipid/fatty acid contents within mouse lungs were quantified using the Oil Red O staining. The protective effect of FAO activator L-carnitine (Lca, 50, 500, or 5 mg/mL) was evaluated by cell counting kit 8 (CCK-8) assay, real-time quantitative PCR (qPCR), ELISA, immunoblotting, fluorescence imaging, and fluorescence plate reader detection in lipopolysaccharide (LPS) (100 ng/mL)-stimulated THP-1-derived macrophages. The in vivo efficacy of Lca (300 mg/kg) was determined in a 10 mg/kg LPS-induced ALI mouse model. We found for the first time that lipid accumulation in pulmonary macrophages was significantly increased in a classical ALI murine model, which indicated disrupted FAO induced by LPS. Lca showed potent anti-inflammatory and antioxidative effects on THP-1 derived macrophages upon LPS stimulation. Mechanistically, Lca was able to maintain FAO, mitochondrial activity, and ameliorate mitochondrial dynamics. In the LPS-induced ALI mouse model, we further discovered that Lca inhibited neutrophilic inflammation and decreased diffuse damage, which might be due to the preservation of mitochondrial homeostasis. These results broadened our understanding of ALI/ARDS pathogenesis and provided a promising drug candidate for this syndrome.

Molecular subtypes based on ferroptosis-related genes and tumor microenvironment infiltration characterization in lung adenocarcinoma.

Recently, several molecular subtypes with different prognosis have been found in lung adenocarcinoma (LUAD). However, the characteristics of the ferroptosis molecular subtypes and the associated tumor microenvironment (TME) cell infiltration have not been fully studied in LUAD. Using 1160 lung adenocarcinoma samples, we explored the molecular subtypes mediated by ferroptosis-related genes, along with the associated TME cell infiltration. The ferroptosis score was constructed using the least absolute shrinkage and selection operator regression (LASSO) method to quantify the ferroptosis characteristics of a single tumor. Three different molecular subtypes related to ferroptosis, with different prognoses, were identified in LUAD. Analysis of TME cell infiltration revealed immune heterogeneity among the three subtypes. Cluster A was characterized by immunosuppression and was associated with stromal activation. Cluster C was characterized by a large number of immune cells infiltrating the TME, promoting tumor immune response, and it was significantly enriched in immune activation-related signaling pathways. Relatively less infiltration of immune cells was a feature of cluster B. The ferroptosis score can predict tumor subtype, immunity and prognosis. A low ferroptosis score was characterized by immune activation and good prognosis, as seen in the cluster C subtype. Relative immunosuppression and poor prognosis were the characteristics of a high ferroptosis score, as seen in cluster A and B subtypes. At the same time, the anti-PD-1/L1 immunotherapy cohort demonstrated that a low ferroptosis score was associated with higher efficacy of immunotherapy. The ferroptosis score is a promising biomarker that could be of great significance to determine the prognosis, molecular subtypes, TME cell infiltration characteristics and immunotherapy effects in patients with LUAD.

GRK6 Depletion Induces HIF Activity in Lung Adenocarcinoma.

G protein-coupled receptor kinase 6 (GRK6) is expressed in various tissues and is involved in the development of several diseases including lung cancer. We previously reported that GRK6 is down-regulated in lung adenocarcinoma patients, which induces cell invasion and metastasis. However, further understanding of the role of GRK6 in lung adenocarcinoma is required. Here we explored the functional consequence of GRK6 inhibition in lung epithelial cells. Analysis of TCGA data was coupled with RNA sequencing (RNA-seq) in alveolar epithelial type II (ATII) cells following depletion of GRK6 with RNA interference (RNAi). Findings were validated in ATII cells followed by tissue microarray analysis. Pathway analysis suggested that one of the Hallmark pathways enriched upon GRK6 inhibition is 'Hallmark_Hypoxia' (FDR = 0.014). We demonstrated that GRK6 depletion induces HIF1α (hypoxia-inducible factor 1 alpha) levels and activity in ATII cells. The findings were further confirmed in lung adenocarcinoma samples, in which GRK6 expression levels negatively and positively correlate with HIF1α expression (P = 0.015) and VHL expression (P < 0.0001), respectively. Mechanistically, we showed the impact of GRK6 on HIF activity could be achieved via regulation of VHL levels. Taken together, targeting the HIF pathway may provide new strategies for therapy in GRK6-depleted lung adenocarcinoma patients.

Clinical value of serum and exhaled breath condensate inflammatory factor IL-11 levels in non-small cell lung cancer: Clinical value of IL-11 in non-small cell lung cancer.

OBJECTIVE: Our study aimed to observe and evaluate the clinical value of interleukin (IL)-11 in the serum and exhaled breath condensate of patients with non-small cell lung cancer (NSCLC). METHODS: A total of 91 patients with NSCLC and 72 healthy volunteers were included in this study. IL-11 concentration was determined by ELISA, and the relationship between IL-11 expression in serum and exhaled breath condensate specimens, and the clinicopathological characteristics of patients with NSCLC were analyzed. The relationship between serum IL-11 expression and traditional tumor markers and inflammation indicators of NSCLC was also analyzed. The correlation between serum IL-11 and exhaled breath condensate IL-11 level was determined. The receiver operating characteristic curve was used to evaluate the diagnostic value of IL-11 and carcinoembryonic antigen single and combined detection for NSCLC. The published data from online databases were used to analyze the relationship between the expression of IL-11 and the prognosis of NSCLC. RESULTS: IL-11 concentration in serum and exhaled breath condensate specimens of patients with NSCLC were significantly increased. IL-11 expression was positively correlated with lymph node metastasis, distant metastasis, tumor node metastasis stage, and tumor differentiation degree of NSCLC. The expression of IL-11 in serum was positively correlated with that in exhaled breath condensate specimens. IL-11 expression was closely related to that of neutrophil-to-lymphocyte ratio and carcinoembryonic antigen. The combination of serum IL-11 with exhaled breath condensate IL-11 and carcinoembryonic antigen showed significantly higher diagnostic value than any one marker alone. Besides, the high IL-11 expression was closely related to the poor prognosis of NSCLC. CONCLUSION: IL-11 can be used as a potential diagnostic and prognostic biomarker for NSCLC.

Epidermal Growth Factor in Exhaled Breath Condensate as Diagnostic Method for Non-Small Cell Lung Cancer.

OBJECTIVE: Lung cancer is one of the most common malignant tumors in humans. Finding a highly sensitive and specific marker is very important. This study investigated the clinical significance of epidermal growth factor in exhaled breath condensate and serum of patients with non-small cell lung cancer. METHODS: From October 17, 2013, to June 5, 2017, exhaled breath condensate and blood samples from 155 patients with non-small cell lung cancer, 63 patients with benign pulmonary nodules, and 115 healthy controls were collected using a breath condenser. Each sample was analyzed by enzyme-linked immunosorbent assay. RESULTS: Epidermal growth factor level in the exhaled breath condensate from the non-small cell lung cancer group (197.86 ± 60.67 pg/mL) was higher than that in the healthy group (124.75 ± 36.09 pg/mL), P < .05. Epidermal growth factor level in the exhaled breath condensate of the smoking group (208.85 ± 40.94 pg/mL) was higher than that of the nonsmoking group (185.52 ± 36.88 pg/mL), P < .05. Epidermal growth factor level in the exhaled breath condensate in phases III and IV of non-small cell lung cancer group (212.17 ± 35.41 pg/mL) was higher than that in phases I and II (173.91 ± 38.08 pg/mL), P < .05. Epidermal growth factor level in the exhaled breath condensate of the death group (241.05 ± 27.19 pg/mL) was higher than that of the survival group (188.75 ± 37.07 pg/mL), P < .05. The epidermal growth factor exhaled breath condensate levels were positively correlated with the serum epidermal growth factor levels with a correlation coefficient of 0.495 (P < .05). The sensitivity and specificity of epidermal growth factor exhaled breath condensate test were 80.0% and 89.6%, respectively. CONCLUSION: The detection of epidermal growth factor level in exhaled breath condensate exhibits is important in the diagnosis, disease monitoring, and prognosis of non-small cell lung cancer.

Hypermethylation of the G protein-coupled receptor kinase 6 (GRK6) promoter inhibits binding of C/EBPα, and GRK6 knockdown promotes cell migration and invasion in lung adenocarcinoma cells.

We previously reported that the expression of G protein-coupled receptor kinase 6 (GRK6) is significantly downregulated in lung adenocarcinoma (LADC) tissues, and low expression levels of GRK6 are correlated with poor survival prognosis. However, the specific regulatory mechanisms and functions of GRK6 in LADC remain unknown. Here, we report that GRK6 mRNA expression levels are downregulated in LADC tissues compared to those in matched adjacent non-tumor tissues (P < 0.001). The promoter of the GRK6 gene was found to be hypermethylated in LADC tissues, and its methylation was correlated with both GRK6 expression and pathology grade. GRK6 promoter hypermethylation may predict shorter overall survival. Treatment with 5-aza-2'-deoxycytidine significantly enhanced GRK6 gene expression. Four binding sites of CCAAT/enhancer-binding protein-α (C/EBPα) in the CpG island of the GRK6 gene promoter were predicted in silico, of which three sites were further confirmed by ChIP. Decreased binding of C/EBPα to binding sites 1, 3 and 4 of the GRK6 gene promoter was observed in LADC tissues. Inhibition of C/EBPα significantly inhibited GRK6 expression, while overexpression of C/EBPα significantly promoted GRK6 expression. In addition, overexpression of GRK6 significantly suppressed, while GRK6 knockdown promoted cell migration and invasion. Overexpression of GRK6 enhanced E-cadherin expression and suppressed vimentin expression, and silencing of GRK6 had the opposite effects. Furthermore, ectopic expression of GRK6 significantly decreased matrix metalloproteinase (MMP) 2 and MMP7 protein expression levels. Our findings suggest that hypermethylation of the GRK6 gene promoter suppressed binding of C/EBPα, thereby contributing to the promotion of cell migration and invasion. The methylation status of the GRK6 promoter might be suitable for use as an epigenetic biomarker, and the C/EBPα-GRK6 signaling pathway may be a potential target for LADC.

Overexpression of fibronectin type III domain containing 3B is correlated with epithelial-mesenchymal transition and predicts poor prognosis in lung adenocarcinoma.

Fibronectin (FN) type III domain containing 3B (FNDC3B), a member of the FN family, regulates the invasion and metastasis of cells in numerous tumor types. However, the mechanisms through which FNDC3B regulates carcinogenesis in lung adenocarcinoma (LADC) tissues have remained elusive. The present study revealed that the protein levels of FNDC3B and vimentin were significantly elevated in LADC tissues compared with those in normal lung tissues. By contrast, the expression of E-cadherin was decreased in LADC tissues compared with that in normal lung tissues. Furthermore, the aberrant expression of FNDC3B and epithelial-mesenchymal transition (EMT) markers was significantly associated with histological differentiation, lymph node metastasis and tumor-nodes-metastasis stage. Kaplan-Meier analysis indicated that a high expression of FNDC3B may be associated with poor overall survival of patients with LADC. In addition, overexpression of FNDC3B promoted the protein expression of EMT-associated genes in the A549 lung adenocarcinoma cell line. In conclusion, the present results support the notion that FNDC3B acts as an oncogene in LADC; it may serve a pivotal role in the development and progression of LADC and may participate in the regulation of the EMT.

Overexpression of RCN1 correlates with poor prognosis and progression in non-small cell lung cancer.

We investigated the expression of reticulocalbin-1 (RCN1) and its prognostic significance in non-small cell lung cancer (NSCLC). RCN1 expression was evaluated by immunohistochemical analysis with tissue microarrays in NSCLC tissues and matched adjacent noncancerous tissues. Furthermore, quantitative polymerase chain reaction and Western blot were also used to examine the expression of RCN1. Moreover, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, clone formation, and transwell assays were used to measure cell proliferation, migration, and invasion. Lastly, we used the Kaplan-Meier method and log-rank test to compare overall survival rates between the RCN1-high expression group and the RCN1-low expression group. In this study, immunohistochemistry by tissue microarray at RCN1 expression was significantly up-regulated in NSCLC tissues compared with adjacent noncancerous tissues. We further confirmed the up-regulation of RCN1 by quantitative polymerase chain reaction and Western blot assay. RCN1 expression level was closely related to lymph node metastasis (P < .001) and TNM stage (P = .012). Kaplan-Meier analysis showed that high RCN1 expression was remarkably associated with poor prognosis with NSCLC patients. A suppression of cell proliferation, migration, and invasion was obtained in A549 cells treated with RCN1 small interfering RNA. Our data indicate that RCN1 expression may have an vital role at promoting the occurrence of NSCLC, and it may be a vital molecular marker in the diagnosis and prognosis of NSCLC patients.

Histone deacetylase 5 promotes the proliferation and invasion of lung cancer cells.

Histone deacetylase 5 (HDAC5), as a member of the class IIa family of HDACs, is frequently dysregulated in human malignancies. However, little is known regarding the specific role of HDAC5 in lung cancer. We aimed to evaluate HDAC5 expression in human lung cancer and to determine the effects of HDAC5 on lung cancer cells. First, the expression levels of both HDAC5 protein and mRNA were evaluated in lung cancer tissues and cell lines by western blot analysis and RT­qPCR, and the results suggested that HDAC5 was significantly upregulated in human lung cancer tissues and cell lines. To address the effects of HDAC5 on the biological behavior of human lung adenocarcinoma cells, we generated human lung cancer A549 cell lines in which HDAC5 was either overexpressed or depleted. The results indicated that overexpression of HDAC5 significantly promoted the proliferation and invasion, and inhibited the apoptosis of A549 cells. On the contrary, HDAC5 knockdown largely decreased the proliferation and invasion and enhanced the apoptosis of A549 cells. Furthermore, we demonstrated that HDAC5 overexpression promoted the expression of DLL4, Six1, Notch 1 and Twist 1 in A549 cells. Downregulation of HDAC5 caused a significant inhibition of the expression of DLL4, Six1, Notch 1 and Twist 1 in A549 cells. Taken together, our data demonstrated that HDAC5 displayed a significant upregulation in lung cancer, and elevated HDAC5 might be involved in the potentiation of proliferation and invasion of lung cancer cells, as well as the inhibition of lung cancer cell apoptosis by the upregulation of DLL4, Six1, Notch 1 and Twist 1. The present study may provide an evidence for the potential application of HDAC5 inhibitors in the therapy of lung cancer.

Speckle-type POZ protein functions as a tumor suppressor in non-small cell lung cancer due to DNA methylation.

BACKGROUND: Tumor suppressor epigenetic silencing plays an important role in non-small cell lung cancer (NSCLC) development and progression. Previously, the expression of speckle-type POZ protein (SPOP) has been found to be significantly inhibited in NSCLC. Our research aimed to investigate the molecular mechanisms, clinical significance and epigenetic alteration of SPOP in NSCLC. MATERIALS AND METHODS: Bisulfite sequencing PCR and methylation-specific PCR were performed to test gene methylation. Chromatin immunoprecipitation (ChIP) was performed to detect transcription factor C/EBPα combinations and the promoter of the SPOP gene. Furthermore, we evaluated the effects of C/EBPα siRNA on SPOP expression, tumor cell migration and proliferation via MTT and Transwell assays in vitro and tumor growth in vivo. The relationship between the methylation status of the SPOP gene and clinicopathologic characteristics was investigated. RESULTS: Hypermethylation was found in the CpG island of the SPOP gene promoter in NSCLC tissues, and this methylation was found to be correlated with SPOP expression. SPOP promoter methylation was associated with the pathology grade. The transcriptional activities were significantly inhibited by the hypermethylation of specific CpG sites within the SPOP gene promoter, while 5-aza-2'-deoxycytidine significantly increased SPOP gene expression. C/EBPα also played a key role in SPOP regulation. Five C/EBPα binding sites in the CpG island of the SPOP gene promoter were identified by ChIP. Inhibition of C/EBPα significantly reduced SPOP expression. SPOP mediated the C/EBPα-regulated suppression of invasion, migration and proliferation in vitro and tumor growth in vivo. CONCLUSIONS: SPOP function and expression in NSCLS were regulated by DNA methylation and C/EBPα transcriptional regulation combination effects, indicating that the SPOP promoter methylation status could be utilized as an epigenetic biomarker and that the C/EBPα-SPOP signaling pathway could be a potential therapeutic target in NSCLC.

Decreased expression of speckle-type POZ protein for the prediction of poor prognosis in patients with non-small cell lung cancer.

Speckle-type POZ domain protein (SPOP) has been acknowledged as a tumor suppressor gene in numerous types of cancer. However, SPOP expression and its prognostic role in human non-small cell lung cancer (NSCLC) remain unknown. The present study investigated SPOP expression in NSCLC and evaluated its prognostic significance in patients with NSCLC. The results demonstrated that SPOP expression was significantly downregulated in NSCLC tissues at the mRNA and protein level compared with normal lung tissues using reverse transcription-quantitative polymerase chain reaction, and western blot analysis. Immunohistochemical staining results also demonstrated that SPOP was expressed at a low level in 84.1% (132/157) of NSCLC samples and at a high level in 52.2% (12/23) of normal lung samples, whereby the difference was statistically significant (P<0.001). In addition, it was revealed that the level of SPOP was associated with histologic type (P=0.003), tumor differentiation (P=0.046), tumor size (P=0.0036), lymph node metastasis (P=0.041) and clinical stages (P=0.046). Furthermore, the overall survival of patients with high SPOP expression was significantly increased compared with that of patients with low SPOP expression (P=0.003). These results revealed that SPOP expression was downregulated in NSCLC tissues and associated with poor prognosis in patients with NSCLC, suggesting that SPOP is an independent prognostic marker candidate for NSCLC.

Effects of Pesticide Mixtures on Zooplankton Assemblages in Aquatic Microcosms Simulating Rice Paddy Fields.

The individual and combined effects of pesticides (chlorpyrifos, triadimefon and butachlor) on the zooplankton assemblages of microcosms were investigated. Laboratory microcosms were constructed with water and sediment to simulate aquatic conditions in China's rice paddy fields. Results from principal response curves analysis showed that butachlor and triadimefon had no significant impact individually on the population level in zooplankton assemblages. The deleterious effects of pesticide mixtures on the zooplankton were mainly caused by chlorpyrifos. In fact, assemblage succession only occurred in the treatments containing chlorpyrifos. There was no synergy effect on the microcosm from combinations of pesticides on the assemblages. The zooplankton assemblages affected by chlorpyrifos did not recover at the termination of the experiment, i.e., after 56 days.

Prognostic value of decreased GRK6 expression in lung adenocarcinoma.

BACKGROUND: In recent years, G protein-coupled receptor kinases (GRKs) have been implicated in cancer metastasis through phosphorylation of the activated form of G protein-coupled receptors. However, little is known of GRK6 expression in lung adenocarcinoma (LADC) and its potential prognostic value in LADC. METHODS: In this study, protein expression of GRK6 was determined in LADC tissues (n = 122) and normal lung tissues (n = 45) by immunohistochemistry (IHC) analysis on tissue microarray (TMA). In addition, mRNA level of GRK6 in matched pairs of cancerous and non-cancerous fresh frozen tissues from 20 LADC patients was investigated using real-time quantitative PCR (qPCR). Furthermore, protein expression level of GRK6 was evaluated in matched pairs of cancerous and non-cancerous fresh frozen tissues from another 18 LADC patients. Univariate and multivariate analyses based on Cox proportional hazards regression models were performed to investigate the correlation between GRK6 expression and overall survival of LADC patients. RESULTS: According to the IHC analysis on TMA, GRK6 expression was significantly down-regulated in LADC patients, but high in normal lung tissue (P < 0.001). Besides, our qPCR and western blot results confirmed GRK6 down-regulation in both mRNA and protein levels in LADC tissues as compared to matched adjacent non-cancerous tissues (all P < 0.001). Additionally, For TMA slides, protein expression of GRK6 was significantly associated with staging (P = 0.030), pathology grade (P = 0.036). Consistent with the associated poor clinicopathologic features, patients with GRK6 low expression tumors had a worse overall survival compared to patients with GRK6 high expression tumors. Further multivariate analysis using the Cox proportional hazards model revealed that GRK6 expression level (P = 0.004) was an independent prognostic factor for overall survival. CONCLUSION: These findings indicate for the first time that decreased expression of GRK6 may serve as an independent predictor of overall survival in LADC patients.

[Effects of sprinkler irrigation on the plant nitrogen accumulation and translocation and kernel protein content of winter wheat].

Taking wheat cultivar Bainong AK58 as test material, a field experiment was conducted to study the plant nitrogen accumulation and translocation and kernel protein content of winter wheat under sprinkler irrigation and surface irrigation, aimed to understand the differences in the nitrogen metabolism characteristics of winter wheat under different irrigation regimes. At booting stage, no significant difference was observed in the total amount of plant nitrogen accumulation between sprinkler irrigation and surface irrigation; while from booting stage to maturing stage, the total amount of plant nitrogen accumulation under sprinkler irrigation was significantly higher. Under sprinkler irrigation, the translocation amount and contribution rate of the nitrogen stored in leaf, glume, stem and sheath at pre-anthesis to the kernel increased, while the contribution rate of the assimilated nitrogen after anthesis to the kernel nitrogen declined. Both the relative protein content and the total protein yield in the kernel increased significantly under sprinkler irrigation. In conclusion, sprinkler irrigation could significantly regulate the nitrogen translocation and kernel protein accumulation of winter wheat.

[Effects of 5-aminolevulinic acid on winter wheat dry matter accumulation after anthesis and flag leaf senescence].

Taking wheat cultivar Bainong AK58 as test material, a field experiment was conducted to study the effects of different concentration 5-aminolevulinic acid (ALA) (0,10, 30 and 50 mg x L(-1)) applied at initial heading stage on the post-anthesis dry matter accumulation and flag leaf senescence of the cultivar. Applying 10-50 mg x L(-1) of ALA benefited the dry matter accumulation, with its total amount at maturing stage being significantly higher than that of the control (0 mg x L(-1) ALA). 10-50 mg x L(-1) of ALA had no significant effects on the distribution of accumulated dry matter in leaf, stem and sheath, and grain, but increased the contribution of the dry matter to grain yield. 10-50 mg x L(-1) of ALA increased the leaf area index at milky and dough stages, but had no effects on it at flowering stage. After treated with 10-50 mg x L(-1) ALA, the leaf SPAD value and net photosynthetic rate from anthesis to milky stage were significantly higher, and the MDA content and relative electric conductivity at later grain-filling stage were lower, compared with those of the control. Applying 10-50 mg x L(-1) of ALA increased the grain number per spike, 1000-grain mass, and grain yield significantly, with the best effect when applying 30 mg x L(-1) ALA.

1-(5-Bromo-2-hydr-oxy-4-methoxy-phen-yl)ethanone.

In the title compound, C(9)H(9)BrO(3), the dihedral angle between the ethanone group and the aromatic ring is 3.6 (2)°. The mol-ecular conformation is consolidated by an intra-molecular O-H⋯O hydrogen bond. The crystal structure is stabilized by π-π inter-actions between the benzene rings [centroid-centroid distance = 3.588 (2) Å].

[Association between residual sleepiness and central sleep apnea events in patients with obstructive sleep apnea syndrome].

OBJECTIVE: To investigate the possible association between residual sleepiness (RS) and central sleep apnea events in patients with obstructive sleep apnea syndrome (OSAS) following continuous positive airway pressure (CPAP) treatment, as well as the effects of adaptive servo-ventilation (ASV) on RS. METHODS: Following correct application of CPAP treatment and exclusion of other sleepiness-associated disorders, 50 patients with moderate-to-severe OSAS were recruited, including 26 patients with RS (RS group) and 24 patients without RS (control group). The treatment of one month's auto-CPAP (AutoCPAP) followed by one week ASV with autoCS2 ventilator was performed. Comparisons were made separately before treatment, on AutoCPAP and ASV treatments in both groups of the following parameters: polysomnographic parameters including central sleep apnea index (CSAI), micro-arousal index (MAI), etc; daytime Epworth sleepiness score (ESS), and possibly sleepiness-associated factor, i.e., plasma tumor necrosis factor-alpha (TNF-alpha). Plasma TNF-alpha levels were measured by enzyme linked immunosorbent assay (ELISA). t test and single factor analysis of variances were used for comparison between two groups and within group respectively. q test was used for couple comparison within group at 3 different stages. Pearson correlation test was performed for correlation analysis between 2 variables. RESULTS: Before treatment there was no significant difference between two groups in apnea hypopnea index (AHI), MAI, minimal pulse oxygen saturation (minSpO2), ESS and plasma TNF-alpha levels (t: 0.630, 1.223, 0.691, 0.764 and 0.19 2, all P > 0.05). However, the CSAI in RS group was significantly higher than that in the control group [(7.19 +/- 1.75) times/h vs (3.37 +/- 1.04) times/h, t = 4.097, P < 0.05)]. After 1 month's AutoCPAP treatment there was a significant decrease in AHI, CSAI, MAI and ESS in both groups (q: 0.87-112.55, all P < 0.05), but CSAI, MAI and ESS in the RS group than those in the control group [CSAI: (7.19 +/- 1.75) times/h vs (3.37 +/- 1.04) times/h, t = 9.473, P < 0.05; MAI: (9.00 +/- 1.95) times/h vs (2.36 +/- 0.66) times/h, t = 14.385, P < 0.05; ESS: 9.54 +/- 0.51 vs 5.42 +/- 1.32, t = 2.857, P < 0.05). On one weeks' ASV treatment there was such a further significant decrease in CSAI, MAI and daytime ESS in the RS group and the control group. In addition, compared with the plasma TNF-alpha level before treatment in the RS group, there was no statistical difference on AutoCPAP treatment but a significant decrease on ASV treatment. Plasma TNF-alpha levels were positively correlated with ESS (r = 0.503, P < 0.01) and MAI (r = 0.545, P < 0.01). CONCLUSIONS: RS in OSAS patients following CPAP treatment was associated with their CSAI before and during treatment. By effectively eliminating CSA events with ASV, RS was significantly improved, which suggested that ASV was effective in treatment of RS. The elevation of plasma TNF-alpha level was correlated with the severity of sleepiness and may be involved in the pathogenesis of RS.

5,3'-Dihydr-oxy-7,4'-dimethoxy-flavanone from Artemisia sphaerocephala Kraschen.

The title compound, C(17)H(16)O(6), was isolated from the Chinese Tibetan medicinal plant Artemisia sphaerocephala Kraschen. The mol-ecular conformation is consolidated by two intra-molecular O-H⋯O hydrogen bonds. A further inter-molecular O-H⋯O hydrogen bond leads to chains along [010] in the crystal structure.

N,N'-Dimeth-oxy-N,N'-dimethyl-succinamide.

The title compound, C(8)H(16)N(2)O(4), is a Weinreb amide that is also an important inter-mediate for the preparation of ketones and aldehydes. The molecule possesses a centre of symmetry.