TAK-242 improves sepsis-associated acute kidney injury in rats by inhibiting the TLR4/NF-κB signaling pathway.

OBJECTIVE: This study was designed to observe the effect of toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) pathway activity on sepsis-associated acute kidney injury (SA-AKI), thereby providing new considerations for the prevention and treatment of SA-AKI. METHODS: The rats were divided into Sham, cecal ligation and puncture (CLP), CLP + vehicle, and CLP + TAK-242 groups. Except the Sham group, a model of CLP-induced sepsis was established in other groups. After 24 h, the indicators related to kidney injury in blood samples were detected. The pathological changes in the kidneys were observed by hematoxylin-eosin staining, and tubular damage was scored. Oxidative stress-related factors, mitochondrial dysfunction-related indicators in each group were measured; the levels of inflammatory factors in serum and kidney tissue of rats were examined. Finally, the expression of proteins related to the TLR4/NF-κB signaling pathway was observed by western blot. RESULTS: Compared with the CLP + vehicle and CLP + TAK-242 groups, the CLP + TAK-242 group reduced blood urea nitrogen (BUN), creatinine (Cr), cystatin-C (Cys-C), reactive oxygen species (ROS), malondialdehyde (MDA), and inflammatory factors levels (p < 0.01), as well as increased superoxide dismutase (SOD) activity of CLP rats (p < 0.01). Additionally, TAK-242 treatment improved the condition of CLP rats that had glomerular and tubular injuries and mitochondrial disorders (p < 0.01). Further mechanism research revealed that TAK-242 can inhibit the TLR4/NF-κB signaling pathway activated by CLP (p < 0.01). Above indicators after TAK-242 treatment were close to those of the Sham group. CONCLUSION: TAK-242 can improve oxidative stress, mitochondrial dysfunction, and inflammatory response by inhibiting the activity of TLR4/NF-κB signaling pathway, thereby preventing rats from SA-AKI.

Predicting the Risk of Acute Kidney Injury in Patients After Percutaneous Coronary Intervention (PCI) or Cardiopulmonary Bypass (CPB) Surgery: Development and Assessment of a Nomogram Prediction Model.

BACKGROUND We sought to create a model that incorporated ultrasound examinations to predict the risk of acute kidney injury (AKI) after percutaneous coronary intervention (PCI) or cardiopulmonary bypass (CPB) surgery. MATERIAL AND METHODS A total of 292 patients with AKI after PCI or CPB surgery were enrolled for the study. Afterwards, treatment-related information, including data pertaining to ultrasound examination, was collected. A random forest model and multivariate logistic regression analysis were then used to establish a predictive model for the risk of AKI. Finally, the predictive quality and clinical utility of the model were assessed using calibration plots, receiver-operating characteristic curve, C-index, and decision curve analysis. RESULTS Predictive factors were screened and the model was established with a C-index of 0.955 in the overall sample set. Additionally, an area under the curve of 0.967 was obtained in the training group. Moreover, decision curve analysis also revealed that the prediction model had good clinical applicability. CONCLUSIONS The prediction model was efficient in predicting the risk of AKI by incorporating ultrasound examinations and a number of factors. Such included operation methods, age, congestive heart failure, body mass index, heart rate, white blood cell count, platelet count, hemoglobin, uric acid, and peak intensity (kidney cortex as well as kidney medulla).

[Effects of Acrolein on the Proliferation of and Per1 Gene Expression in Pulmonary Epithelial Cells].

OBJECTIVE: To investigate the effect of acrolein on the proliferation of pulmonary epithelial cells and its possible mechanism. METHODS: Two strains of pulmonary epithelial cells, A549 cells and MLE15 cells, were used as in vitro models of pulmonary epithelial cell, and were treated with 80 µmol/L acrolein or phosphate buffer saline (PBS) as the control. The proliferation of pulmonary epithelial cells were determined with CCK-8 kit after cell culturing resumed for 12 h, 24 h, 36 h and 48 h post acrolein treatment, and the expression of period circadian regulator gene 1 ( Per1) was examined using Western blot test 24 h after acrolein treatment. In addition, after acrolein treatment, the cells were restored with transforming growth factor-ß (TGF-ß) added in the medium, and the cell proliferation and the expression of Per1 protein were also examined. RESULTS: The proliferation of A549 cells and MLE15 cells decreased significantly after being treated with 80 µmol/L acrolein for 30 min, and the expression of Per1 protein was also downregulated significantly ( P<0.05). The addition of TGF-ß after acrolein treatment did not significantly change the reduction in cell proliferation caused by acrolein, but the expression of Per1 protein in pulmonary epithelial cells was significantly higher than that in cells restored without TGF-ß ( P<0.05). CONCLUSION: Acrolein treatment resulted in the decreased proliferation of pulmonary epithelial cells and the Per1 expression in pulmonary epithelial cells. Although TGF-ß addition did not reverse the reduction of cell proliferation after acrolein treatment, the Per1 expression levels were recovered to a certain extent compared to that in cells restored in medium without TGF-ß after acrolein treatment.

[Dietary Patterns and Their Association with Diabetes Mellitus in Middle-Aged and Older Rural Population in Xinxiang County, Henan Province].

OBJECTIVE: To analyze the prevalence of diabetes mellitus among middle-aged and older rural adults of Xinxiang county, Henan Province and its correlation with dietary patterns. METHODS: The study was done based on the data collected from a cross-sectional survey of Xinxiang County, which was part of the Prospective Cohort Study on the Common Chronic Non-Communicable Diseases in Rural areas of Henan Province. Randomized cluster sampling was used to select adult respondents (≥18 years old) from among the residents of 17 villages in Xinxiang county. The respondents completed questionnaires, and underwent physical examinations and laboratory tests between April, 2017 and June, 2017. A total of 7604 individuals aged between 45 and 79 were included in our study. Dietary patterns were established through factor analysis and the dietary pattern factor scores were divided into quartiles (Q1-Q4). The relationship between dietary patterns and diabetes mellitus was analyzed with multivariate logistic regression model. RESULTS: Out of the total of 7604 middle-aged and older rural adults in Xinxiang County, Henan Province, 1604 had diabetes mellitus, suggesting a 21.1% prevalence of diabetes mellitus. Factor analysis was used to establish four dietary patterns, namely animal-based diet, vegetable-egg diet, mixed diet and traditional diet. Subjects of these four dietary patterns displayed different demographic characteristics. There were no statistical difference in anthropometricor clinical indicators between the quartile with the lowest dietary pattern factor score (Q1) and the quartile with the highest dietary pattern factor score (Q4) for subjects with animal-based diet ( P>0.05). Compared with those in the Q1 quartile of vegetable-egg diet, subjects in the Q4 quartile of vegetable-egg diet showed lower levels of high-density lipoprotein cholesterol (HDL-C), along with different distribution of fasting blood glucose (FBG), showing statistically significant difference ( P<0.05). In comparison to subjects in Q1 quartile of mixed diet, those in Q4 quartile showed lower levels of systolic blood pressure (SBP), the difference being statistically significant ( P<0.05). In the traditional diet group, subjects in the Q4 quartile had lower waist circumference (WC), but higher levels of HDL-C than those of subjects in Q1 quartile. In addition, the distribution of glycated-hemoglobin (HbA1c) and FBG were different, the difference being statistically significant ( P<0.05). The results of multivariate logistic regression analysis demonstrated that traditional diet could be a protective factor of diabetes mellitus (odds ratio [ OR]=0.810, 95% CI: 0.690-0.952, P trend<0.05) after adjusting for multiple confounding factors. CONCLUSION: The prevalence of diabetes in middle-aged and older rural residents is relatively high in Xinxiang County, Henan Province, and there may be a protective relationship between traditional diet and diabetes mellitus.

[Renal Injury and Its Mechanisms in Lepr db/ db Mice].

OBJECTIVE: To study the mechanism of renal injury in Lepr db/ db mice with the leptin receptor homozygous deficiency. METHODS: Ten male of 28-week-old Lepr db/+ mice with leptin receptor heterozygous deficiency were selected as control group and ten male Lepr db/ db mice with leptin receptor homozygous deficiency were used in this study. After fasting for 8 hours, the body mass, fasting blood glucose (FBG) and glycosylated hemoglobulin (HbA1c) of the mice were measured. Blood of the mice was obtained from femoral artery before euthanasia. Serum creatinine (CRE), blood urea nitrogen (BUN), superoxide dismutase (SOD), glutathione (GSH) and malonaldehyde (MDA) were detected by corresponding kits, and serum interleukin-1ß (IL-1ß), monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α) were measured using enzyme-linked immunosorbent assay (ELISA) method. The kidney was taken for pathological observation. The expression levels of nuclear factor E2-related factor 2 (Nrf2) and nuclear factor kappa B (NF-κB) in renal were analyzed by Western blot. The mitochondria of renal was isolated by the corresponding kit. Meanwhile, the expression level of lipoic acid synthase (LIAS) in renal mitochondria was measured by Western blot. RESULTS: The body mass, FPG, HbA1c, CRE and BUN levels of the Lepr db/ db mice were significantly increased in comparison with the Lepr db/+ mice ( P<0.05). Compared with the Lepr db/+ mice, the Lepr db/ db mice renal exhibited glomerular hypertrophy, thickened basement membrane and capillary wall, the mesangial matrix expansion and mesangial cell hyperplasia. Compared with the Lepr db/+ mice, the serum level of GSH in the Lepr db/ db mice was decreased significantly ( P<0.05). The levels of MDA and concentrations of MCP-1, IL-1ß and TNF-α in serum of the Lepr db/ db mice were higher than those of the Lepr db/+ mice ( P<0.05). Compared with the Lepr db/+ mice, the expression of LIAS and Nrf2 protein in the Lepr db/ db mice renal were decreased ( P<0.05), while the expression of NF-κB protein was increased ( P<0.05). CONCLUSION: LIAS, Nrf2 and NF-κB might play significant roles through regulation of oxidative stress and inflammation in the renal injury of Lepr db/ db mice.

LncRNA H19 and Target Gene-mediated Cleft Palate Induced by TCDD.

This study investigated the role of long non-coding RNAs (lncRNAs) in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression levels of long non-coding RNA H19 (lncRNA H19) and insulin-like growth factor 2 (IGF2) gene were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lncRNA H19 and IGF2 gene specifically showed embryo age-related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lncRNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lncRNA H19 and IGF2 can mediate the development of mouse cleft palate.

Meta-analysis comparing the safety and efficacy of metastatic colorectal cancer treatment regimens, capecitabine plus irinotecan (CAPIRI) and 5-fluorouracil/leucovorin plus irinotecan (FOLFIRI).

The relative efficacy and safety of first-line metastatic colorectal cancer (mCRC) treatment regimens, capecitabine with irinotecan (CAPIRI) and 5-fluorouracil/leucovorin plus irinotecan (FOLFIRI), are not well defined. We identified and subsequently examined seven independent, randomized controlled clinical trials, performing a meta-analysis to compare these two treatment regimens. Using Medline, EMBASE, Cochrane Library (CENTRAL), and the American Society of Clinical Oncology Annual Meeting to search available literature until February 2014, we identified seven studies comparing safety and efficacy of CAPIRI and FOLFIRI in mCRC patients. These studies were pooled and evaluated for rates of progression-free survival (PFS), objective response rate (ORR), overall survival (OS), and diarrhea. CAPIRI and FOLFIRI demonstrated similar efficacy outcomes, though CAPIRI was associated with a higher incidence of diarrhea. CAPIRI and FOLFIRI are equally effective options for first-line treatment of mCRC.

The expression profiles of microRNAs in Kaposi's sarcoma.

Kaposi's sarcoma (KS) is a multicentric angioproliferative tumor of mesenchymal origin. The molecular and biologic aspects of KS are not fully understood. MicroRNAs are non-protein-coding small RNAs in the size range 19-25 nucleotides (nt) that play important roles in biological processes, including cellular differentiation, proliferation, and death. We performed a miRNA microarray analysis by detecting six paired KS and matched adjacent healthy tissues using the 7th generation of miRCURY(TM) LNA Array (v.18.0) (Exiqon) containing 3100 capture probes. We selected 10 significant differentially expressed miRNAs, which were confirmed by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) in 18 paired KS and matched adjacent healthy tissue specimens. We also investigated the associations between clinical features and miRNA expression. Among the 3100 human miRNA probes in the microarrays, we identified 170 differentially expressed miRNAs (69 upregulated and 101 downregulated miRNAs) in KS versus adjacent healthy tissues. Among the most significantly upregulated miRNAs were miR-126-3p, miR-199a-3p, miR-16-5p, and the 13 KSHV-related miRNAs. The most significantly downregulated miRNAs included miR-125b-1-3p and miR-1183. Eight upregulated miRNAs, miR-181b-5p, miR-199a-3p, miR-15a-5p, miR-126-3p, miR-1297, kshv-miR-k12-12-3p, kshv-miR-k12-1-5p, and miR-16-5p, and two downregulated miRNAs, miR-125b-1-3p and miR-1183, were confirmed by qRT-PCR in 18 paired KS samples. The qRT-PCR results for 10 miRNAs were consistent with our microarray results. The miR-125b-1-3p and miR-16-5p had statistically significant associations with HHV-8 and HIV infections in KS. The results of miRNA profiling showed that KS appears to have unique expression patterns when compared with paired adjacent healthy tissues, suggesting that deregulation of miRNAs plays an important role in the progression of KS. These differentially expressed miRNAs may provide novel diagnostic and prognostic tools.

FBXL5 targets cortactin for ubiquitination-mediated destruction to regulate gastric cancer cell migration.

Cortactin, an actin-interacting protein, is implicated in cytoskeletal architecture and often amplified in several types of cancer including gastric adenocarcinomas. Downregulation of cortactin decreases cell migration and invasion. However, how to regulate cortactin in gastric cancer remains largely unknown. Here, we report that FBXL5 interacts with and targets cortactin for ubiquitylation and subsequent proteasomal degradation. Furthermore, we showed that FBXL5-induced cortactin degradation is mediated by extracellular regulated signal kinase (ERK). Serine phosphorylation sites mutant, cortactinS405A/S418A, prevent FBXL5-induced cortactin degradation. Moreover, CortactinS405A/S418A exhibited stronger effects in promoting gastric cancer cell migration when compared to wild-type cortactin. Taken together, our data suggested a novel molecular mechanism for the negative regulation of cortactin by FBXL5 in gastric cancer cells migration.

[Inductive effect of zinc oxide nanoparticles on interleukin 8 gene expression in human bronchial epithelial cells and its regulatory mechanism].

OBJECTIVE: To clarify the effect of zinc oxide nanoparticles (ZnO-NPs) (30 nm in diameter) on the interleukin 8 (IL-8) gene expression in human bronchial epithelial cells (BEAS-2B) and its regulatory mechanism. METHODS: BEAS-2B cells were used in the study. The MTT assay was employed to evaluate the damage to BEAS-2B cells by ZnO-NPs. RT-PCR and ELISA were used to measure the mRNA and protein expression levels of IL-8 in the BEAS-2B cells exposed to ZnO-NPs. The IL-8 mRNA decay assay was used to determine the effect of ZnO-NPs on IL-8 mRNA stability. RESULTS: Exposure to ZnO-NPs significantly increased the level of IL-8 mRNA in BEAS-2B cells and the level of IL-8 protein in supernatant medium. The transcription inhibitor significantly reduced the mRNA expression of IL-8 induced by ZnO-NPs. ZnO-NPs significantly delayed IL-8 mRNA degradation in the BEAS-2B cells that were pretreated with actinomycin D for terminating IL-8 mRNA synthesis. CONCLUSION: ZnO-NPs can increase the mRNA and protein expression levels of IL-8 and IL-8 mRNA stability in BEAS-2B cells.

[Coal tar pitch smoke extract-induced pyroptosis in human bronchial epithelial cells].

OBJECTIVE: To explore whether coal tar pitch smoke extract (CTP) induced pyroptosis in human bronchial epithelial cells (BEAS-2B). METHODS: BEAS-2B cells were treated with different concentrations of CTP (1, 3 µg/ml) for 8h and 24 h, respectively. Lactic dehydrogenase (LDH) activity and interleukin-1 beta (IL-1ß) levels in the supernatants of cell culture media were measured with LDH activity or human IL-1ß ELISA kit, respectively. The activity of Caspase-1 was measured with Caspase-1 colorimetric assay kit. RESULTS: The activity of caspase-1 in 1 and 3 µg/ml CTP groups were (9.29 ± 0.30) and (8.67 ± 0.59) µmol/ml respectively which were both significantly increased compared to that (7.42 ± 0.59) µmol/ml in the control group (P < 0.05) after 8 h exposure, but there was no significant difference in the activity of LDH and levels of IL-1ß in the cell culture media among the CTP and control groups. 24 h after exposure, the activity of LDH in the CTP (1, 3 µg/ml) groups were (1323.03 ± 28.53) and (1148.45 ± 16.42) U/dl respectively which were significantly higher than that (1091.93 ± 26.64) U/dl in the control group (P < 0.05), and the levels of IL-1ß in the CTP (1 and 3 µg/ml) groups were (125.37 ± 25.00) pg/ml and (92.04 ± 19.09) pg/ml respectively which were significantly higher than that (46.20 ± 14.43) pg/ml in the control group (P < 0.05), but there was no significant difference in the activity of Caspase-1 among CTP and control groups (P < 0.05). CONCLUSION: CTP treatment induced early increase in caspase-1 activity followed by the increase in LDH activity and IL-1 levels, indicative of pyroptosis in human bronchial epithelial cells.

Association between serum sodium level trajectories and survival in patients with heart failure.

AIMS: The effect of changes in serum sodium levels on the survival of patients with heart failure (HF) is unclear. We aimed to analyse the impact of serum sodium level trajectories on survival in intensive care unit (ICU) patients with HF. METHODS: A total of 4760 patients diagnosed with HF between 2001 and 2012 from the Medical Information Mart for Intensive Care III (MIMIC-III) database were extracted. Of these patients, 1132 patients who died within 48 h of ICU admission were excluded, and 3628 patients were included in this retrospective cohort study. Sodium levels were measured at baseline, 6, 12, 18, 24, 30, 36, 42, and 48 h. Patients were divided into hyponatremia, normal, and hypernatremia groups based on baseline sodium levels, and trajectory modelling was performed for each group separately. Group-based trajectory model (GBTM) method was utilized to identify serum sodium levels trajectories. RESULTS: The number of patients with hyponatremia (<135 mmol/L), normal sodium levels (135-145 mmol/L), and hypernatremia (>145 mmol/L) at baseline were 594 (16.37%), 2,738 (75.47%), and 296 (8.16%), respectively. A total of seven trajectory groups were identified, including hyponatremia-slow rise group [initial levels (IL), 128.48 ± 5.42 mmol/L; end levels (EL), 131.23 ± 3.83 mmol/L], hyponatremia-rapid rise to normal group (IL, 132.13 ± 2.18 mmol/L; EL, 137.46 ± 3.68 mmol/L), normal-slow decline group (IL, 137.65 ± 2.15 mmol/L; EL, 134.50 ± 2.54 mmol/L), normal-steady-state group (IL, 139.20 ± 2.26 mmol/L; EL, 139.04 ± 2.58 mmol/L), normal-slow rise group (IL, 140.94 ± 2.37 mmol/L; EL, 143.43 ± 2.89 mmol/L), hypernatremia-rapid decline to normal group (IL, 146.31 ± 1.98 mmol/L; EL, 140.71 ± 3.61 mmol/L), and hypernatremia-slow decline group (IL, 148.89 ± 5.54 mmol/L; EL, 146.28 ± 3.90 mmol/L). The results showed that hyponatremia-slow rise group [hazard ratio (HR) = 1.35; 95% confidence interval (CI), 1.01-1.80, P = 0.040], hyponatremia-rapid rise to normal group (HR = 1.37; 95% CI, 1.11-1.71, P = 0.004), hypernatremia-rapid decline to normal group (HR = 1.46; 95% CI, 1.08-1.97, P = 0.014), and hypernatremia-slow decline group (HR = 1.49; 95% CI, 1.07-2.07, P = 0.018) trajectories were associated with an increased risk of 1-year mortality in HF patients compared with normal-steady-state group. After adjustment for all confounders, hyponatremia-rapid rise to normal group (HR = 1.26, 95% CI; 1.01-1.57, P = 0.038) and hypernatremia-rapid decline to normal group (HR = 1.36; 95% CI, 1.01-1.84, P = 0.047) trajectories were still related to an increased risk of 1-year mortality in patients with HF. CONCLUSIONS: Serum sodium level trajectories were associated with mortality in patients with HF. Association between serum sodium level trajectories and prognosis in patients with HF deserve further study.

Combined exposure of heat stress and ozone enhanced cognitive impairment via neuroinflammation and blood brain barrier disruption in male rats.

BACKGROUND: Heat stress (HS) exposure has been linked to cognitive dysfunction. In reality, high temperature does not occur alone in environment, and ozone (O3) and heatwaves usually co-exist in atmospheric environment. However, whether O3 exposure exacerbates HS-induced cognitive impairment and the potential underlying mechanisms have not been explored experimentally. The aim of this study was to determine the co-effects and mechanisms of HS and O3 on the cognitive dysfunction. METHODS: 48 Sprague Dawley male rats were randomly divided into 4 groups: control, HS, O3 and HS plus O3 (HO3) groups. Rats in HS and HO3 group were exposed to 40 °C every morning from 9:00 to 12:00 for 15 consecutive days. While rats in O3 and HO3 groups were exposed to 0.7 ppm O3 the same day from 14:00 to 17:00 for 15 days. Cognitive performance was examined with Morris water maze test. Neurodegeneration, glial activation, neuroinflammation, blood brain barrier (BBB) disruption and apoptosis were evaluated by Western blot, Elisa, immunohistochemistry and immunofluorescence staining. RESULTS: HS induced cognitive decline and neuronal damage in rats. Further studies showed that exposure of rats to HS could also induce glial activation, neuroinflammation and neuronal apoptosis in hippocampus, and decrease in the expressions of ZO-1, claudin-5 and occluding, indicative of BBB disruption. Impressively, the neuronal effects induced by HS, as depicted above, could be worsened by co-exposure to O3 in rats. CONCLUSIONS: Co-exposure to O3 promotes HS-induced cognitive impairment in rats possibly through glial-mediated neuroinflammation and BBB disruption.

[Theoretical study of the influence of different substituents on the electron-photon spectra of quinoline].

Different substituents result in different changes in electron-photon spectra, and to reveal the relationship between substituents and spectra, a theoretical investigation was elaborated via quantum chemical calculations. Density functional theory and single excitation configuration interaction were respectively employed in optimizing geometric and electronic structures of ground and excited states, and the absorption and emission spectra were studied by time-dependent density functional theory methods. The results show that all the different substituents bring on different geometric and electronic structures of ground and excited states, different energies of frontier molecular orbitals as well as different pi-conjugated systems, the spectra change with all the differences, and relationships are brought out in this paper, which gives theoretical reference for identifying different derivatives from electron-photon spectra.

[Effects of MICT/HIIT on the ultrastructure of myocardium and soleus in rats with high-fat diet and its mechanisms].

Objective: To investigate the effects of moderate intensity continuous training (MICT) and high intensity intermittent training (HIIT) on the ultrastructure of myocardium and soleus in rats with high fat diet, and to explore the mechanisms. Methods: 5-week-old male SD rats were randomly divided into normal diet quiet group (C), high-fat diet quiet group (F), high-fat MICT group (M) and high-fat HIIT group (H), with 8 rats in each group, and the fat content of the high-fat dietary feed was 45%. The M and H groups were given 12 weeks of treadmill running with an incline of 25°. The M group was given continuous exercise with 70%VO2max intensity, and the H group was given intermittent exercise with 5 min 40%~45%VO2max and 4 min 95%~99%VO2max intensity successively. After the intervention, the contents of free fatty acid (FFA), triglyceride (TG), high density lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (LDL) in serum were detected. Transmission electron microscopy was used to observe the ultrastructure of myocardium and soleus in rats. Western blot was used to detect the protein expressions of AMPK, malonyl-CoA decarboxylase (MCD) and carnitine palmitoyl transterase 1 (CPT-1) in myocardium and soleus. Results: Compared with C group, the body weight, Lee's index, the contents of LDL, TG and FFA in serum were increased, the content of HDL was decreased (P＜0.05), the protein expressions of AMPK and CPT-1 in myocardium and soleus were increased, the protein expression of MCD was decreased (P＜0.05), and the ultrastructure was damaged in group F. Compared with F group, the body weight and Lee's index were decreased, the contents of LDL and FFA in serum were decreased (P＜0.01), the protein expressions of AMPK, MCD and CPT-1 in myocardium were increased, and the protein expressions of AMPK and MCD in soleus were increased (P＜0.05), and the ultrastructural damage was attenuated in M and H groups. Compared with M group, the content of HDL in serum was increased (P＜0.01), the protein expressions of AMPK and MCD in myocardium were increased, and the ultrastructural damage was mild, the protein expression of AMPK in soleus was decreased, the protein expression of MCD in soleus was increased (P＜0.05), and the ultrastructural damage was severe in group H. Conclusion: MICT and HIIT have different effects on the ultrastructure of myocardium and soleus in high-fat diet rats by intervening the protein expression of AMPK, MCD and CPT-1.

[The change of telomere protein in BEAS-2B malignant transformation cell induced by coal tar pitch smoke extracts].

OBJECTIVE: By testing the changes of telomere binding protein in malignant transformation BEAS-2B cells induced by coal tar pitch smoke extracts, to study the role of protection of telomeres 1 (POT1), telomeric repeat binding factor 1 (TRF1) and TRF2 in tumorgenesis that contact with coal tar pitch. METHODS: The BEAS-2B cells were induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, the protein expression variations were determined by cell culture overslip of immunohistochemical methods. RESULTS: In malignant transformation cells, the mRNA expression level (POT1: 0.63 ± 0.04, TRF1: 0.36 ± 0.01) and the protein expression level (POT1: 0.36 ± 0.05, TRF1: 0.09 ± 0.03) of POT1 and TRF1 was statistically significant decreased compared to that of BEAS-2B group (mRNA: POT1: 1.00 ± 0.04, TRF1: 1.01 ± 0.16; protein: POT1: 0.55 ± 0.07, TRF1: 0.27 ± 0.07) and DMSO group (mRNA: POT1: 0.89 ± 0.12, TRF1: 0.90 ± 0.08; protein: POT1: 0.55 ± 0.10, TRF1: 0.26 ± 0.04) (P < 0.05); mRNA expression level (1.45 ± 0.07) and the protein expression level (0.88 ± 0.06) of TRF2 was increased compared to that of BEAS-2B group (mRNA: 1.00 ± 0.07, protein: 0.48 ± 0.06) and DMSO group (mRNA: 1.00 ± 0.06, protein: 0.50 ± 0.06) (P < 0.05). CONCLUSION: The change of gene and protein expression level in POT1, TRF1, and TRF2 involved in the process that evolved into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.

Microscopic and Macroscopic Fragmentation Characteristics under Hypervelocity Impact Based on MD and SPH Method.

This work investigates the difference in the fragmentation characteristics between the microscopic and macroscopic scales under hypervelocity impact, with the simulations of Molecular Dynamics (MD) and Smoothed Particle Hydrodynamics (SPH) method. Under low shock intensity, the model at microscopic scale exhibits good penetration resistance due to the constraint of strength and surface tension. The bullet is finally embedded into the target, rather than forming a typical debris cloud at macroscopic scale. Under high shock intensity, the occurrence of unloading melting of the sample reduces the strength of the material. The material at the microscopic scale has also been completely penetrated. However, the width of the ejecta veil and external bubble of the debris cloud are narrower. In addition, the residual velocity of bullet, crater diameter and expansion angle of the debris cloud at microscopic scale are all smaller than those at macroscopic scale, especially for low-velocity conditions. The difference can be as much as two times. These characteristics indicate that the degree of conversion of kinetic energy to internal energy at the microscopic scale is much higher than that of the macroscopic results. Furthermore, the MD simulation method can further provide details of the physical characteristics at the micro-scale. As the shock intensity increases, the local melting phenomenon becomes more pronounced, accompanied by a decrease in dislocation atoms and a corresponding increase in disordered atoms. In addition, the fraction of disordered atoms is found to increase exponentially with the increasing incident kinetic energy.

[Change of structural maintenance of chromosome (SMC)1, SMC3, Separase and Securin expression in BEAS-2B malignant transformation cell induced by coal tar pitch smoke extracts].

OBJECTIVE: to study the role of structural maintenance of chromosome (SMC)1, SMC3, Separase and Securin in tumorgenesis that contact with coal tar pitch. METHODS: the BEAS-2B cells was induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, and the protein expression variation were determined by cell culture overslip of immunohistochemical methods. RESULTS: in malignant transformation cells, the mRNA and the protein expression level of SMC1 gene was not statistically significantly different compared with the BEAS-2B group and DMSO group (P > 0.05); SMC3 and Separase was increased and Securin was decreased (P < 0.05), while the difference between other two control groups was not significant (P > 0.05). CONCLUSIONS: the up expression level of SMC3 and Separase and the down expression level of Securin are involved in the process that evolves into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.

[Construction of lentiviral mediated CyPA siRNA and its functions in non-small cell lung cancer].

OBJECTIVE: To construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer. METHODS: First, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM. RESULTS: It was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05). CONCLUSION: Lentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.

[Involvement of Toll-like receptor in silica-induced tumor necrosis factor alpha release from human macrophage cell line].

OBJECTIVE: To characterize the role of Toll-like receptor 4 (TLR4) in silica-induced production of tumor necrosis factor alpha (TNFalpha) from macrophage cell line. METHODS: The human macrophage cell line THP-1 was incubated with silica suspension. Cell media were collected and TNFalpha levels in the supernatants measured with ELISA. To examine the involvement of TLR4 in silica-induced TNFalpha release, the neutralizing antibody (HTA125) against human TLR4 receptor was employed to pretreat THP-1 cells prior to silica treatment. Moreover, murine macrophages expressing wild type or mutated TLR4 were also treated with silica to verify the effect of TLR4 in silica-induced TNFalpha release. RESULTS: Compared with the control group [(3.18 +/- 0.41) pg/ml], the TNFalpha release in cells exposed to 100 microg/ml silica for 4 h and 8 h [(4.71 +/- 0.84), (6.22 +/- 0.58) pg/ml, respectively] increased 1.48 and 1.96 fold, respectively. Pretreatment of THP-1 cells with 20 microg/ml HTA125 antibody significantly blocked silica-induced TNFalpha release by 27%. Furthermore, the TNFalpha content released from cells expressing mutated TLR4 reduced by 30% in compared with that from the cells expressing wild type TLR4 after silica stimulation. CONCLUSION: TLR4 mediates silica-induced TNFalpha release from macrophages.