Yes-associated protein regulates cortical actin architecture and dynamics through intracellular translocation of Rho GTPase-activating protein 18.

Yes-associated protein (YAP) is a transcriptional co-activator that controls the transcription of target genes and modulates the structures of various cytoskeletal architecture as mechanical responses. Although it has been known that YAP regulates actin-regulatory proteins, the detailed molecular mechanism of how they control and coordinate intracellular actin architecture remains elusive. Herein, we aimed to examine the structure and dynamics of intracellular actin architecture from molecular to cellular scales in normal and YAP-knockout (YAP-KO) cells utilizing high-speed atomic force microscopy (HS-AFM) for live-cell imaging and other microscope-based mechanical manipulation and measurement techniques. YAP-KO Madin-Darby canine kidney cells had a higher density and turnover of actin filaments in the cell cortex and a higher elastic modulus. Laser aberration assay demonstrated that YAP-KO cells were more resistant to damage than normal cells. We also found that Rho GTPase-activating protein 18 (ARHGAP18), a downstream factor of YAP, translocated from the cortex to the edge of sparsely cultured YAP-KO cells. It resulted in high RhoA activity and promotion of actin polymerization in the cell cortex and their reductions at the edge. HS-AFM imaging of live cell edge and a cell-migration assay demonstrated lower membrane dynamics and motility of YAP-KO cells than those of normal cells, suggesting lower actin dynamics at the edge. Together, these results demonstrate that a YAP-dependent pathway changes the intracellular distribution of RhoGAP and modulates actin dynamics in different parts of the cell, providing a mechanistic insight into how a mechano-sensitive transcription cofactor regulates multiple intracellular actin architecture and coordinates mechano-responses.

[Cerebellar ferroptosis of hypertension mice following lead exposure].

OBJECTIVE: Exploring the changes in cerebellar ferroptosis in hypertensive mice after lead exposure. METHODS: Twenty-five healthy C57 male mice were selected to construct a hypertensive model by intraperitoneal injection of angiotensin II(Ang II) at a concentration of 0.05 mg/kg for 7 consecutive days. After a systolic blood pressure of 140 mmHg, 20 hypertensive mice were randomly divided into a hypertensive control group and a hypertensive lead exposure group. Twenty C57 mice with normal blood pressure were randomly divided into a blood pressure normal control group and a blood pressure normal lead exposure group. The mice in the normal blood pressure control group and the hypertensive control group drank water freely. Mice in the lead exposure group with normal blood pressure and the lead exposure group with hypertension drank lead acetate water containing 250 mg/L. Ang II was injected intraperitoneally every two days in the hypertensive control group and hypertensive lead exposed group mice. Each group of mice was poisoned for 12 weeks. Using open field experiments and balance beam experiments to detect motor dysfunction in mice. Using a reagent kit to detect the levels of divalent iron(Fe~(2+)), malondialdehyde(MDA), and glutathione(GSH) in the cerebellum of different groups of mice. Western blot was used to determine the protein expression of member 11 of the solute carrier family 7(SLC7A11), glutathione peroxidase 4(GPX4), nuclear receptor coactivator 4(NCOA4), microtubule associated protein 1 light chain 3B(LC3B), and ferritin heavy chain 1(FTH1) in mouse cerebellar tissue. RESULTS: The result of the open field experiment showed that the activity distance(1013.04 cm) of mice in the hypertensive lead exposure group was significantly lower than that of the hypertensive control group(1351.18 cm) and the lead exposure group with normal blood pressure(1287.35 cm). And the lead exposure group with hypertension also extended the time through the balance beam, which was 29.40 seconds(P<0.05). In addition, the Fe~(2+)content in the cerebellum of mice in the hypertensive lead exposure group was 3.33 µmol/g prot, which was 1.54 times that of the hypertensive control group and 1.14 times that of the lead exposure group with normal blood pressure. The MDA content was 4.71 nmol/mg prot, higher than that of the hypertensive control group and the lead exposure group with normal blood pressure. The GSH content was 5.36 µmol/g prot, lower than that of the hypertensive control group and the lead exposure group with normal blood pressure(P<0.05). Western blot result showed that compared with the hypertensive control group and the lead exposure group with normal blood pressure, the protein expression of SLC7A11 and GPX4 in the hypertensive lead exposure group was significantly reduced(P<0.05). In addition, compared with the control group with normal blood pressure, the expression of NCOA4 and LC3B proteins in the cerebellum of mice in the hypertension control group and lead exposure group with normal blood pressure increased, while the expression of FTH1 protein decreased(P<0.05). The expression of NCOA4 and LC3B proteins in the hypertensive lead exposure group was higher than that in the hypertensive control group and the lead exposure group with normal blood pressure, while the expression of FTH1 protein decreased(P<0.05). CONCLUSION: Lead exposure can exacerbate iron death in the cerebellar tissue of hypertensive mice, and iron autophagy may be involved in its occurrence and development.

EBV promotes vascular mimicry of dormant cancer cells by potentiating stemness and EMT.

Vascular mimicry (VM) is defined as a vascular channel-like structure composed of tumor cells that correlates with the growth of cancer cells by providing blood circulation. However, whether VM can be formed in dormant cancer cells remains unclear. Our previous research revealed that polyploid giant cancer cells (PGCCs) are specific dormant cells related to the poor prognosis of head and neck cancer. Here, we demonstrated that EBV could promote VM formation by PGCCs in vivo and in vitro. Furthermore, we revealed that the activation of the ERK pathway partly mediated by LMP2A is responsible for stemness, and the acquisition of the stemness phenotype is crucial to the malignant biological behavior of PGCCs. The epithelial-to-mesenchymal transition (EMT) process plays a considerable role in PGCCs, and EMT progression is vital for EBV-positive PGCCs to form VM. This is the first study to reveal that EBV creates plasticity in PGCC-VM and provide a new strategy for targeted anti-tumor therapy.

[Neurobehavioral damage and Th17 cell infiltration in mice exposed to nano carbon black].

OBJECTIVE: The effects of nano-carbon black on neural behavior and Th17 cell infiltration in mice were investigated by establishing a mice model of subacute dynamic inhalation of carbon black aerosol. METHODS: 36 SPF grade male C57BL/6 mice were randomly divided into a control group(clean air), a low carbon black group(15 mg/m~3), and a high carbon black group(30 mg/m~3). Nano-carbon black particles were blown into the dynamic exposure cabinet by aerosol generator for 28 days. Morris water maze test and open field test were used to detect the neural behavior of mice. The pathological changes of prefrontal cortex in mice were observed by HE staining. The proportion of Th17/CD4~+ cells in peripheral blood and brain tissue of mice was detected by flow cytometry. Western blotting was used to detect the protein expression of interleukin(IL)-17 and IL-23 in the prefrontal cortex of mice. RESULTS: The result of open field test showed that compared with the control group, the central area residence time and standing times of mice in the low and high carbon black groups decreased significantly(P<0.05), and the defecation times of mice in the high carbon black group increased significantly(P<0.05). The central area residence time of mice in the high carbon black group was significantly lower than that in the low carbon black group(P<0.05). The Morris water maze result showed that the escape latency of the high carbon black group mice on the 3rd day was significantly higher than that of the control group(P<0.05). Meanwhile, the escape latency of the carbon black group mice on the 4th day was significantly higher than that of the control group(P<0.05). The positioning navigation test showed that the number of mice crossing the platform in the high carbon black group was significantly higher than that in the control group(P<0.05). The HE staining result showed that the neural cells in the prefrontal cortex of the control group mice were round, the cytoplasm was plump and evenly distributed, and the nucleus was clearly visible in an oval shape. The low carbon black group showed that the neural cells were deep staining of nerve cells, blurred structure, and nuclear pyknosis. The high carbon black group further intensified. The flow cytometry result showed that compared with the control group, the percentage of Th17/CD4~+T cells in the peripheral blood of the carbon black group mice was significantly increased, and the high carbon black group mice were significantly higher than the low carbon black group(P<0.05). Meanwhile, the percentage of Th17/CD4~+T cells in the brain tissue of carbon black treated mice significantly increased(P<0.05). The high carbon black group was significantly higher than the low carbon black group(P<0.05). Western blotting result showed that compared with the control group, the expression of IL-17 and IL-23 proteins in the prefrontal cortex of the carbon black group mice brain tissue was significantly increased(P<0.05). Compared with the low carbon black group, the expression of IL-17 and IL-23 proteins in the prefrontal cortex of the high carbon black group mice brain tissue was significantly increased(P<0.05). The difference was statistically significant. CONCLUSION: Nano-carbon black exposure can lead to an increase in Th17 cells in peripheral blood and brain tissue of mice, which in turn promotes damage to the prefrontal cortex of mice, and ultimately causes neurobehavioral changes in mice.

Norepinephrine promotes glioma cell migration through up-regulating the expression of Twist1.

BACKGROUND: Glioma cells are characterized by high migration ability, resulting in aggressive growth of the tumors and poor prognosis of patients. It has been reported that the stress-induced hormone norepinephrine (NE) contributes to tumor progression through mediating a number of important biological processes in various cancers. However, the role of NE in the regulation of glioma migration is still unclear. Epithelial-to-mesenchymal transition (EMT) is one of the most important steps for tumor migration and metastasis. Twist1, as a key regulator of EMT, has been found to be elevated during glioma migration. But it is still unknown whether Twist1 is involved in the effect of NE on the migration of glioma cells. METHODS: Wound healing assay and transwell assay were conducted to evaluate the migration of glioma cells upon different treatments. The mesenchymal-like phenotype and the expression of Twist1 after NE treatment were assessed by cell diameters, real-time PCR, western blot and immunofluorescence staining. The gain-and loss-of-function experiments were carried out to investigate the biological function of Twist1 in the migration induced by NE. Finally, the clinical significance of Twist1 was explored among three public glioma datasets. RESULTS: In this study, our finding revealed a facilitative effect of NE on glioma cell migration in a ß-adrenergic receptor (ADRB)-dependent way. Mechanistically, NE induced mesenchymal-like phenotype and the expression of Twist1. Twist1 overexpression promoted glioma cells migration, while knockdown of Twist1 abolished the discrepancy in the migration ability between NE treated glioma cells and control cells. In addition, the clinical analysis demonstrated that Twist1 was up-regulated in malignant gliomas and recurrent gliomas, and predicted a poor prognosis of glioma patients. CONCLUSIONS: NE enhanced the migration ability of glioma cells through elevating the expression of Twist1. Our finding may provide potential therapeutic target for protecting patients with glioma from the detrimental effects of stress biology on the tumor progression.

MiR-378a-3p/ SLC7A11 regulate ferroptosis in nerve injury induced by lead exposure.

An increasing number of studies have clarified that ferroptosis plays a vital role in neurodegenerative diseases, which is characterized by the accumulation of Fe2+, lipid peroxidation, and alteration of mitochondrial structure. However, whether ferroptosis is involved in nerve injury caused by lead exposure remains unclear. In this study, HT22 cells and mice were treated with lead acetate to investigate the role of ferroptosis in lead neurotoxicity. The results showed that lead exposure resulted in an accumulation of Fe2+, an increase in malondialdehyde (MDA) levels, and a decrease in glutathione (GSH) levels in vivo and in vitro. An increase in the levels of lipid reactive oxygen species (ROS) and the expression of 4HNE, as well as the change in mitochondrial morphology, were also observed in HT22 cells treated with lead acetate. In addition, deferoxamine (DFO; an iron chelator) attenuated the accumulation of Fe2+ and significantly enhanced the viability of HT22 cells exposed to lead. Fer-1 (an anti-ferroptosis agent) reduced the level of lipid ROS and expression of 4HNE in lead-treated HT22 cells. Furthermore, lead exposure sharply downregulated the expression of SLC7A11 in HT22 cells. Overexpression of SLC7A11 reversed the changes in MDA and GSH levels and cell viability induced by lead exposure. In contrast, lower expression of SLC7A11 accelerated the changes in these parameters. Consequently, we screened miRNAs that regulate SLC7A11 using TargetScan. We found that miR-378a-3p showed the highest expression among the target miRNAs regulating SLC7A11 expression. Inhibition of miR-378a-3p expression reversed the reduction in GSH and the increase in lipid ROS levels induced by lead exposure. Taken together, these findings indicate that lead exposure can cause ferroptosis and that miR-378a-3p exerted an important effect by regulating SLC7A11 expression. Our findings provide new insights into the mechanisms underlying the effects of lead exposure.

A prospective study on the difference of clinical outcomes between elderly and adult patients with allergic rhinitis.

OBJECTIVE: The guiding significance of existing guidelines for the diagnosis, treatment and health management of AR in elderly patients is unclear. The aim of this study is to analyze the clinical characteristics and therapeutic effects of elderly and adult AR patients by prospective study. METHODS: A total of 131 AR patients were recruited and divided into elderly group and adult group according to age. After receiving the same pharmacological treatment for 4 weeks, the differences of the two groups in clinical scores including TNSS-4, RQLQ and VAS were compared. RESULTS: After 4 weeks treatment, all clinical scores in the adult group were improved compared with the baseline levels, while in the elderly group, only the TNSS-4 score was significantly reduced, and the RQLQ and VAS scores were not significantly improved. The changes of TNSS-4, RQLQ, and VAS scores in the elderly group were significantly inferior to those in the adult group (LS mean differences were 1.60, 8.80, and 11.10, respectively; P < 0.001). CONCLUSION: We confirmed that elderly and adult AR patients had different clinical characteristics and outcomes, and the degree of improvement in the adult group was significantly better than that in the elderly group. Therefore, it is urgent for us to establish a clinical guideline suitable for the elderly AR population to give more scientific and reasonable recommendations for diagnosis and treatment.

[Role of ferroptosis in cerebellar injury of mice following lead exposure].

OBJECTIVE: To investigate the role of ferroptosis in cerebellar injury of mice following lead exposure. METHODS: A total of forty SPF C57 mice were randomly divided into control group, low-dose lead exposure group, middle-dose lead exposure group and high-dose lead exposure group, with 10 mice in each group. Mice in three lead exposure groups were given 0.25, 0.50, 1.00 g/L lead acetate through drinking water for twelve weeks respectively. Lead concentration was detected by inductively coupled plasma mass spectrometer. The motor function was detected by beam walking test and open field test. Pathological changes of cerebellum in mice were observed by H&E staining. Western blotting was used to detect the protein expression of transferrin receptor-1(TFR-1), ferroportin(FPN-1), solute carrier family 7 member 11(SLC7 A11), glutathione peroxidase 4(GPX4), NF-E2-related factor 2(Nrf2) and heme oxygenase-1(HO-1). RESULTS: The lead concentration in cerebellum of mice in low lead group, medium lead group and high lead group were(1.05±0.11), (1.21±0.10) and(1.48±0.1) µg/g, respectively, which were significantly higher than that in the control group. The time to traverse the beam in low lead group, medium lead group and high lead group was 1.34, 1.64 and 2.02 folds of that in control group, respectively. Open field test showed that the central residence time and standing times of mice in low lead group, medium lead group and high lead group were significantly lower than that in control. Purkinje cells in the cerebellum of mice exposed to different doses of lead showed irregular arrangement, small cell bodies and deep staining, especially in the high lead group. The relative levels of iron in low lead group, medium lead group and high lead group was 1.77, 2.29 and 3.77 folds of that in control group, respectively. The content of MDA in cerebellum of mice in three lead exposure groups increased significantly, while the GHS decreased significantly. Compared with the control group, the expression of TFR-1 protein increased significantly in the lead exposure group, while the expression of FPN-1 protein decreased significantly only in the medium lead group and high lead group, which was 60% and 50% of the control group. Compared with the control group, the expressions of oxidative stress regulatory proteins SLC7 A11 and GPX4 in medium lead group and high lead group decreased significantly. Lead exposure significantly decreased the expression of Nrf2 and HO-1 protein in cerebellum, especially in high lead group. CONCLUSION: In this experiment condition, lead may induce ferroptosis in cerebellum of mice, of which, Nrf2/HO-1 signaling pathway might be involved in, and then further result in motor dysfunction of mice.

A cross-sectional survey of preschool children: Exploring heavy metal exposure, neurotransmitters, and neurobehavioural relationships and mediation effects.

BACKGROUND: Exposure to heavy metals has been considered harmful and can cause cognitive deficits in preschool children. OBJECTIVE: To investigate the possible mediation effect of neurotransmitters on the relationship of heavy metal exposure with neurobehaviour. METHODS: The levels of blood heavy metals and neurotransmitters, along with the neurobehavioural scores, were determined in preschool children. Multiple linear regression was used to assess the relationship between heavy metals, neurotransmitters, and neurobehavioural scores. Furthermore, the mediating role of neurotransmitters was investigated. RESULTS: An interquartile range (IQR) increase in lead (6.10 µg/L) was associated with a decrease of 8.52%, 30.06%, and 20.10% for Glutamic acid (Glu), Glycine (Gly), and gamma-aminobutyric acid (GABA), respectively. An IQR increase in arsenic (19.37 µg/L) was associated with an increase of 6.32% and 2.09% for Gly and GABA, respectively. Further, an IQR increase in zinc (15.58 µg/L) was associated with an increase of 1.44% for Ser, whereas the IQR increase was associated with a decrease of 2.14%, 2.24%, and 1.89% for Glu, Gly, and GABA, respectively. An IQR increase in selenium (38.75 µg/L) was associated with an increase of 1.88% for GABA. Moreover, both Glu and Gly decreased by 2.87% for an IQR increase in manganese (16.92 µg/L). An IQR increase in mercury (15.22 µg/L) was associated with a decrease of 2.43% for Ser, but the IQR increase was associated with an increase of 4.99% and 3.09% for Gly and GABA, respectively. It was found that Glu and Serine (Ser) have a significant linear relationship with conduct score and impulsivity-hyperactivity index, and that there was a significant linear relationship between Ser and the learning disability index. GABA and conduct score and attention-deficit hyperactivity disorder (ADHD) index have a significant linear relationship. There is a significant linear relationship between Gly and conduct, anxiety, ADHD, and impulsivity-hyperactivity index. The results of the mediating effect analysis indicated that Ser, Glu, Gly, and GABA have a specific mediating effect between blood heavy metals and neurobehaviour. CONCLUSION: We showed the mediating effect of neurotransmitters. The current study may provide valuable information regarding the prevention and management of metal-related neurological disorders in preschool children.

High-fat diet exacerbates lead-induced blood-brain barrier disruption by disrupting tight junction integrity.

Environmental exposure to lead (Pb) can damage to the central nervous system (CNS) in humans. High-fat diet (HFD) also has been suggested to impair neurocognitive function. Blood-brain barrier (BBB) is a rigorous permeability barrier for maintaining homeostasis of CNS. The damage of BBB caused by tight junctions (TJs) disruption is central to the etiology of various CNS disorders. This study aimed to investigate whether HFD could exacerbate Pb exposure induced the destruction of BBB integrity by TJs disruption. To this end, we measured cell viability assay, trans-endothelial electrical resistance assay, horseradish peroxidase flux measurement, Western blot analysis, and immunofluorescence experiments. The results showed that palmitic acid (PA), the most common saturated fatty acid found in the human body, can increase the permeability of the BBB in vitro which formed in bEnd.3 cells induced by Pb exposure, and decrease the expression of TJs, such as zonula occludins-1 (ZO-1) and occludin. Besides, we found that PA could promote the up-regulation of matrix metalloproteinase (MMP)-9 expression and activate the c-Jun N-terminal kinase (JNK) pathway induced by Pb. MMP-9 inhibitor or JNK inhibitor could increase BBB integrity and up-regulate the expressions of ZO-1 and occludin after treatment, respectively. Moreover, the JNK inhibitor could down-regulate the expression of MMP-9. In conclusion, these results suggested that HFD exacerbates Pb-induced BBB disruption by disrupting TJs integrity. This may be because PA promotes the activation of JNK pathway and then up-regulated the expression of MMP-9 after Pb-exposure. It is suggested that people with HFD exposed to environmental Pb may cause more serious damage to the CNS.

Effects of lead exposure on the activation of microglia in mice fed with high-fat diets.

Lead (Pb) exposure can cause central nervous system (CNS) damage. The process of Pb neurotoxicity is accompanied by the microglia activation. In addition, microglia activation was observed under the intervention of high-fat diets (HFD). This study was designed to investigate the effect of Pb on the cognitive function of mice with HFD, with focus on the microglia activation in brain. Male C57BL/6J mice, 8 weeks of age, were randomly divided into control, HFD, Pb, and HFD + Pb groups. The results showed that HFD following Pb exposure could exacerbate the learning and memory impairment in mice. Pb exposure could promote microglia activation and increase the expression of M1 microglia marker and decrease the expression of M2 microglia marker in the hippocampus of mice with HFD. Our finding suggested that Pb exposure may aggravate CNS damage by promoting M1 polarization and inhibiting M2 polarization of hippocampal microglia in HFD mice.

A composite prepared from gold nanoparticles and a metal organic framework (type MOF-74) for determination of 4-nitrothiophenol by surface-enhanced Raman spectroscopy.

Core-shell nanoparticles (NPs) consisting of a gold core and a metal-organic framework shell (type MOF-74) were synthesized via one-pot synthesis. The NPs exhibit highly sensitive and stable SERS activity for the detection of 4-nitrothiophenol, with a specific band at 1337 cm-1. The method has a linear response in 0.10-10 µmol·L-1 analyte concentration range and a lower detection limit of 69 nmol·L-1. The potential application of this novel SERS substrate was evaluated by two model reactions involving 4-nitrothiophenol. The first involves in-situ SERS monitoring of the surface plasmon-induced nitration of aromatic rings without adding conventional acid catalyst. The second involves the photocatalytic reduction of 4-nitrothiophenol to 4-thioaminophenol in the presence of Au/MOF-74 under 785-nm laser irradiation. The plasmon-assisted dimerization of 4-nitrothiophenol to form 4,4'-dimercaptoazobenzene can also be monitored simultaneously. Graphical abstract Schematic presentation of a nanoparticle SERS substrate consisting of gold core and MOF-74 shell, which was applied to detection of 4-nitrothiophenol. The Au/MOF-74 was successfully used for in-situ monitoring of two model reactions involving 4-nitrothiophenol by SERS.

Enzyme Inhibitor Antibiotics and Antibiotic-Associated Diarrhea in Critically Ill Patients.

BACKGROUND This study aimed to analyze the factors associated with the development of antibiotic-associated diarrhea (AAD) in critically ill patients receiving enzyme inhibitor antibiotics. MATERIAL AND METHODS A retrospective study of patients with and without AAD admitted to the intensive care unit (ICU) of the First Teaching Hospital of Xi'an Jiaotong University from February 1, 2014, to January 31, 2016, was undertaken. Relevant clinical data underwent univariate or multivariate regression analysis. RESULTS Of 184 patients who received enzyme inhibitor antibiotic therapy, 70 patients (38.04%) developed AAD, with a mean duration of onset of 6.97±3.64 days. AAD was associated with the use of enzyme inhibitor antibiotic therapy alone (OR, 1.142; 95% CI, 1.038-1.256; P=0.007), and in combination with antifungal agents (OR, 2.449; 95% CI, 1.116-5.372; P=0.025), quinolones (OR, 5.219; 95% CI, 1.746-15.601; P=0.003), and oxazolidinones (OR 2.895; 95% CI, 1.183-7.083; P=0.020). The mean duration of ICU stay was significantly increased in patients with AAD (19.00±11.49 days vs. 9.60±6.76 days) (P<0.001). Mean duration of antibiotic therapy (14.09±8.82 days vs. 8.10±4.91 days) (P<0.001) and duration of enzyme inhibitor antibiotic therapy (9.26±5.06 days vs. 6.61±3.24 days) (P<0.001) were significantly increased in patients with AAD. CONCLUSIONS Duration of use of enzyme inhibitor antibiotic therapy and the combined use of antifungals, quinolones, and oxazolidinones increased the incidence and duration of AAD and increased the length of stay in ICU.

[Impairment of the hypothalamus and striatum of rats following lead exposure].

OBJECTIVE: To investigate the impairment of lead exposure on rat hypothalamus and striatum. METHODS: Forty-five healthy SPF male SD rats were randomly divided into three groups, control group( double distilled water), low( 60 mg/kg BW) and high dose( 120 mg/kg BW) groups. The rats were treated with lead acelate water solution in lead exposure group by gavage for 9 weeks. Open-field test was used to detect behavioral changes of the rats. The lead content of hypothalamus and striatum was determined by inductively coupled plasma mass spectrometry( ICP-MS). The mRNA expressions of toll-like receptor 4( TLR4), nuclear transcription factor kappa B( NF-κB), tumor necrosis factors alpha( TNF-α), interleukin-1( IL-1ß) and 8-oxoguanine DNA glycosylase( OGG1) were measured by real-time PCR. Enzyme linked immunosorbent assay( ELISA) was applied to detect the protein content of TLR4, NF-κB, TNF-α, IL-1ß and 8-hydroxy-2-deoxyguanosine( 8-OHd G). RESULTS: There were no significant differences in terms of body weight among three groups of rats. Compared with the control group, total movement distance, the total number of lattice and the distance of the central region in low and high lead exposure group were significantly decreased( P < 0. 05). The lead contents in hypothalamus and striatum of rats of low and high lead exposure group were( 60. 10 ± 6. 71), ( 71. 20 ± 11. 24), ( 44. 07 ± 9. 63)and( 66. 67 ± 8. 78) µg/g, respectively, higher than those in the control group(( 33. 77± 8. 19), ( 25. 75 ± 6. 33) µg/g)( P < 0. 05). While compared with the control group, the mRNA expression of NF-κB and OGG1 of the low and high lead exposure group were( 3. 47 ± 0. 15), ( 1. 43 ± 0. 16) and( 0. 67 ± 0. 13), ( 0. 57 ± 0. 19) folds in hypothalamus, there were marked differences in the mRNA expression of TLR4 and NF-κB between low and high lead group. The NF-κB, TNF-α, IL-1ß protein contents in hypothalamus of the low lead exposure group were( 5. 85 ± 1. 10), ( 56. 15 ± 6. 96) and( 1. 18 ± 0. 20) ng/g, respectively higher than those in control group(( 3. 03 ± 0. 71), ( 49. 25 ± 7. 21) and( 0. 86 ± 0. 11) ng/g)( P < 0. 05). The TLR4, NF-κB, TNF-α, IL-1ß protein contents in hypothalamus of the high lead exposure group were( 0. 67 ± 0. 12), ( 4. 74 ± 0. 68), ( 69. 73 ± 9. 61) and( 1. 43 ± 0. 29) ng/g, respectively, higher than those in control group. There were marked differences in the protein contents of TLR4 and TNF-α significant between low and high lead group in hypothalamus. The mRNA expression of TLR4, NF-κB, IL-1ß in the striatum of rats in the high lead exposure group were significantly higher than those in control group( P < 0. 05). The TLR4, NF-κB, TNF-α, IL-1ß, 8-OHd G protein contents of the high lead exposure group were( 0. 33 ±0. 02) ng/g, ( 4. 66 ± 0. 51), ( 82. 63 ± 7. 99), ( 1. 92 ± 0. 35) and( 1. 21 ± 0. 14) ng/g, respectively higher than those in control group( P < 0. 05), NF-κB protein content of the high lead exposure group was higher than that in low lead exposure group( P < 0. 05). CONCLUSION: Lead exposure does result in the impairment of hypothalamus and striatum, indicating that inflammation and oxidative damage might be involved in this process.

[Gas chromatography for determination of N-isopropylaniline in workplace atmosphere].

OBJECTIVE: To establish a method for determination of N-isopropylaniline in the workplace atmosphere by gas chromatography. METHODS: Air samples were collected by silica gel tube and desorbed by acetone. Then they were separated through DB-WAX columns and N-isopropylaniline was determined by flame ionization detector. RESULTS: The concentration of N-isopropylaniline showed a good linear relationship within the range of 1.40∼665.0 µg/ml. The sampling efficiency was 100%. The accuracy was 96%∼ 99% and the precision was 2.1%∼7.0%. The minimum detectable concentration was 0.056 mg/m(3) (with sampled air volume of 7.5 L). CONCLUSION: The method meets the requirements of analysis and applies to the determination of N-isopropylaniline in the workplace atmosphere.

[Effects of nano-lead exposure on learning and memory as well as iron homeostasis in brain of offspring rats].

OBJECTIVE: To investigate the effects of nano-lead exposure on learning and memory and iron homeostasis in the brain of the offspring rats on postnatal day 21 (PND21) and postnatal day 42 (PND42). METHODS: Twenty adult pregnant female Sprague-Dawley rats were randomly divided into control group and nano-lead group. Rats in the nano-lead group were orally administrated 10 mg/kg nano-lead, while rats in the control group were administrated an equal volume of normal saline until PND21. On PND21, the offspring rats were weaned and given the same treatment as the pregnant rats until 42 days after birth. The learning and memory ability of offspring rats on PND21 and PND42 was evaluated by Morris water maze test. The hippocampus and cortex s amples of offspring rats on PND21 and PND42 were collected to determine iron and lead levels in the hippocampus and cortex by inductively coupled plasma-mass spectrometry. The distributions of iron in the hippocampus and cortex were observed by Perl's iron staining. The expression levels of ferritin, ferroportin 1 (FPN1), hephaestin (HP), and ceruloplasmin (CP) were measured by enzyme-linked immunosorbent assay. RESULTS: After nano-lead exposure, the iron content in the cortex of offspring rats on PND21 and PND42 in the nano-lead group was significantly higher than those in the control group (32.63 ± 6.03 µg/g vs 27.04 ± 5.82 µg/g, P<0.05; 46.20 ±10.60 µg/g vs 36.61 ± 10.2µg/g, P<0.05). The iron content in the hippocampus of offspring rats on PND42 in the nano-lead group was significantly higher than that in the control group (56.9 ± 4.37µg/g vs 37.71 ± 6.92µg/g, P<0.05). The Perl's staining showed massive iron deposition in the cortex and hippocampus in the nano-lead group. FPNl level in the cotfex of offspring rats on PND21 in the nano-lead group was significantly lower than that in the control group (3.64 ± 0.23 ng/g vs 4.99 ± 0.95 ng/g, P<0.05). FPN1 level in the hippocampus of offspring rats on PND42 in the nano-lead group was significantly lower than that in the control group (2.28 ± 0.51 ng/g vs 3.69 ± 0.69 ng/g, P<0.05). The escape latencies of offspring rats on PND21 and PND42 in the nano-lead group were longer than those in the control group (15.54 ± 2.89 s vs 9.01 ± 4.66 s; 6.16 ± 1.42 s vs 4.26 ± 1.51 s). The numbers of platform crossings of offspring rats on PND21 and PND42 in the nano- lead group were significantly lower than those in the control group (7.77 ± 2.16 times vs 11.2 ± 1.61 times, P<0.05; 8.12 ± 1.51 times vs 13.0 ± 2.21 times, P<0.05). ONCLUSION: n Nano-lead exposure can result in iron homeostasis disorders in the hippocampus and cortex of offspring rats and affect their learning and memory ability.

Efficacy, immunogenicity and safety of the HPV-16/18 AS04-adjuvanted vaccine in healthy Chinese women aged 18-25 years: results from a randomized controlled trial.

This phase II/III, double-blind, randomized trial assessed the efficacy, immunogenicity and safety of the human papillomavirus (HPV)-16/18 AS04-adjuvanted vaccine in young Chinese women (ClinicalTrials.gov registration NCT00779766). Women aged 18-25 years from Jiangsu province were randomized (1:1) to receive HPV vaccine (n = 3,026) or Al(OH)3 control (n = 3,025) at months 0, 1 and 6. The primary objective was vaccine efficacy (VE) against HPV-16/18 associated 6-month persistent infection (PI) and/or cervical intraepithelial neoplasia (CIN) 1+. Secondary objectives were VE against virological and clinical endpoints associated with HPV-16/18 and with high-risk HPV types, immunogenicity and safety. Mean follow-up for the according-to-protocol cohort for efficacy (ATP-E) was ∼15 months after the third dose. In the ATP-E (vaccine = 2,889; control = 2,894), for initially HPV DNA negative and seronegative subjects, HPV-16/18 related VE (95% CI) was 94.2% (62.7, 99.9) against 6-month PI and/or CIN1+ and 93.8% (60.2, 99.9) against cytological abnormalities. VE against HPV-16/18 associated CIN1+ and CIN2+ was 100% (-50.4, 100) and 100% (-140.2, 100), respectively (no cases in the vaccine group and 4 CIN1+ and 3 CIN2+ cases in the control group). At Month 7, at least 99.7% of initially seronegative vaccine recipients had seroconverted for HPV-16/18; geometric mean antibody titres (95% CI) were 6,996 (6,212 to 7,880) EU/mL for anti-HPV-16 and 3,309 (2,942 to 3,723) EU/mL for anti-HPV-18. Safety outcomes between groups were generally similar. The HPV-16/18 AS04-adjuvanted vaccine is effective, immunogenic and has a clinically acceptable safety profile in young Chinese women. Prophylactic HPV vaccination has the potential to substantially reduce the burden of cervical cancer in China.

[Study of quercetin on pulmonary fibrosis by silica particles].

OBJECTIVE: To observe the intervention effect of quercetin to silica dust cause pulmonary fibrosis. METHODS: Forty-eight healthy male adult SPF SD rats were selected and they were randomly divided into six groups (control group, 7 d,14 d,21 d and 28d dust group, preventive group). Rats in the control group were administrated 1 ml saline via trachea injection. Rats in dust group and preventive group were give silica solution for 1ml at dose of 50 mg/ml, and the prevention group with quercetin of 50 mg/kg every day lavage treatment intervention. In building on days 7,14,21,28 later, the lung tissue were removaled to HE staining for determining the degree of alveolitis and pulmonary fibrosis. The content of hydroxyproline (HYP) and the activities of catalase (CAT) and glutathione peroxidase (GSH-Px) were detected by using the kits. RESULTS: Alveolar septal edema, inflammatory cell infiltration and fibrosis were not found in control group. In the preventive group, lots of inflammatory cells infiltration were observed on days 7. Inflammatory cells were reduced, the number of the fibroblasts and matrix in alveolar septum were obviously increased, and alveolar structure was damaged on day 14. Pulmonary fibrosis was increased, severe fibrosis was found on day 28. Silicon dust after infected lung tissue expression of HYP content increased, the activity of CAT and GSH-Px decrease gradually. After joining quercetin the expression of HYP content gradually reduce, CAT and GSH-Px activity increased, the difference was statistically significant (P<0.05). CONCLUSION: Quercetin of silica dust caused by pulmonary fibrosis have certain prevention.

[Use of THP-1 for allergens identification method validation].

OBJECTIVE: Look for an in vitro test method to evaluate sensitization using THP-1 cells by the changes of the expression of cytokines to provide more reliable markers of the identification of sensitization. METHOD: The monocyte-like THP-1 cells were induced and differentiated into THP-1-macrophages with PMA (0.1 microg/ml). The changes of expression of cytokines at different time points after the cells being treated with five known allergens, 2,4-dinitrochlorobenzene (DNCB), nickel sulfate (NiSO4), phenylene diamine (PPDA) potassium dichromate (K2Cr2O7) and toluene diisocyanate (TDI) and two non-allergens sodium dodecyl sulfate (SDS) and isopropanol (IPA) at various concentrations were evaluated. The IL-6 and TNF-alpha production was measured by ELISA. The secretion of IL-1beta and IL-8 was analyzed by Cytometric Bead Array (CBA). RESULTS: The section of the IL-6, TNF-alpha, IL-1beta and IL-8 were the highest when THP-1 cells were exposed to NiSO4, DNCB and K2Cr2O7 for 6h, PPDA and TDI for 12h. The production of IL-6 were approximately 40, 25, 20, 50 and 50 times for five kinds chemical allergens NiSO4, DNCB, K2Cr2O7, PPDA and TDI respectively at the optimum time points and the optimal concentration compared to the control group. The expression of TNF-alpha were 20, 12, 20, 8 and 5 times more than the control group respectively. IL-1beta secretion were 30, 60, 25, 30 and 45 times respectively compared to the control group. The production of IL-8 were approximately 15, 12, 15, 12 and 7 times respectively compared to the control group. Both non-allergens SDS and IPA significantly induced IL-6 secretion in a dose-dependent manner however SDS cause a higher production levels, approximately 20 times of the control. Therefore IL-6 may not be a reliable marker for identification of allergens. TNF-alpha, IL-1beta and IL-8 expressions did not change significantly after exposed to the two non-allergens. CONCLUSION: The test method using THP-1 cells by detecting the productions of cytokines (TNF-alpha, IL-1beta and IL-8) can effectively distinguish chemical allergens and non-allergens. The three cytokines may be reliable markers for the identification of potential sensitizing chemicals.