A highly sensitive and rapid LC-MS/MS method for quantification of bexarotene in mouse plasma and brain tissue: Application to mice pharmacokinetic study.

Emerging evidence has suggested that bexarotene, a nearly 20-year-old skin cancer drug, may be a potential drug candidate to treat Alzheimer's disease (AD) and other neurodegenerative disorders. As described in this study, a highly sensitive and rapid method, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine bexarotene in mouse plasma and brain tissue, was established and validated for the first time. Single-step protein precipitation utilizing methanol solution (containing 0.05 % acetic acid) as precipitation agent was employed to prepare the samples of plasma and brain tissue. Chromatographic separation in gradient elution mode was conducted via an Agilent ZORBAX SB-C18 column (50 mm × 4.6 mm, 5 µm) employing methanol-ammonium acetate buffer (5 mM, pH adjusted to 4.6 with acetic acid) as mobile phase which flowed at 0.45 mL/min. The total run time was 6 min for each sample. Detection through mass spectrometric technique was operated by selected reaction monitoring (SRM) in negative electrospray ionization mode. The method was linear within the range of 10.0-15000 ng/mL for plasma and 10.0-600 ng/mL for brain tissue homogenate with the lower limit of quantification of 10.0 ng/ml. The plasma or tissue homogenate was only required 20 µL. The intra- and inter-day precision were less than 13.8 %, and the RE was between -7.4 % and 3.4 %. The method was applied to investigate the plasma pharmacokinetics and brain distribution of bexarotene in mice after being intragastrically administered with bexarotene at the dosage of 100 mg/kg. The results showed that both brain and plasma concentrations of bexarotene peaked at 1.0 h. Bexarotene was rapidly eliminated with a half-life of 2.0 h.

YouTube Videos Related to the Fukushima Nuclear Disaster: Content Analysis.

BACKGROUND: YouTube (Alphabet Incorporated) has become the most popular video-sharing platform in the world. The Fukushima Daiichi Nuclear Power Plant (FDNPP) disaster resulted in public anxiety toward nuclear power and radiation worldwide. YouTube is an important source of information about the FDNPP disaster for the world. OBJECTIVE: This study's objectives were to examine the characteristics of YouTube videos related to the FDNPP disaster, analyze the content and comments of videos with a quantitative method, and determine which features contribute to making a video popular with audiences. This study is the first to examine FDNPP disaster-related videos on YouTube. METHODS: We searched for the term "Fukushima nuclear disaster" on YouTube on November 2, 2019. The first 60 eligible videos in the relevance, upload date, view count, and rating categories were recorded. Videos that were irrelevant, were non-English, had inappropriate words, were machine synthesized, and were <3 minutes long were excluded. In total, 111 videos met the inclusion criteria. Parameters of the videos, including the number of subscribers, length, the number of days since the video was uploaded, region, video popularity (views, views/day, likes, likes/day, dislikes, dislikes/day, comments, comments/day), the tone of the videos, the top ten comments, affiliation, whether Japanese people participated in the video, whether the video recorder visited Fukushima, whether the video contained theoretical knowledge, and whether the video contained information about the recent situation in Fukushima, were recorded. By using criteria for content and technical design, two evaluators scored videos and grouped them into the useful (score: 11-14), slightly useful (score: 6-10), and useless (score: 0-5) video categories. RESULTS: Of the 111 videos, 43 (38.7%) videos were useful, 43 (38.7%) were slightly useful, and 25 (22.5%) were useless. Useful videos had good visual and aural effects, provided vivid information on the Fukushima disaster, and had a mean score of 12 (SD 0.9). Useful videos had more views per day (P<.001), likes per day (P<.001), and comments per day (P=.02) than useless and slightly useful videos. The popularity of videos had a significant correlation with clear sounds (likes/day: P=.001; comments/day: P=.02), vivid information (likes/day: P<.001; comments/day: P=.007), understanding content (likes/day: P=.001; comments/day: P=.04). There was no significant difference in likes per day (P=.72) and comments per day (P=.11) between negative and neutral- and mixed-tone videos. Videos about the recent situation in Fukushima had more likes and comments per day. Video recorders who personally visited Fukushima Prefecture had more subscribers and received more views and likes. CONCLUSIONS: The possible features that made videos popular to the public included video quality, videos made in Fukushima, and information on the recent situation in Fukushima. During risk communication on new forms of media, health institutes should increase publicity and be more approachable to resonate with international audiences.

Long noncoding RNA CBR3 antisense RNA 1 promotes the aggressive phenotypes of non­small­cell lung cancer by sponging microRNA­509­3p and competitively upregulating HDAC9 expression.

Long noncoding RNA CBR3 antisense RNA 1 (CBR3­AS1) plays significant roles in the initiation and progression of osteosarcoma. The aim of the present study was to investigate the involvement of CBR3­AS1 in the development of non­small cell lung cancer (NSCLC). Reverse transcription­quantitative PCR was performed to detect CBR3­AS1 expression in NSCLC tissues and cell lines. The impacts of CBR3­AS1 on cellular proliferation, apoptosis, migration and invasiveness in vitro, and tumor growth in vivo, were investigated using the Cell Counting Kit­8 assay, flow cytometry, Transwell migration and invasion assays, and tumor xenograft model­based analysis, respectively. The results indicated that CBR3­AS1 was markedly upregulated in NSCLC tissues and cell lines. High CBR3­AS1 expression was correlated with larger tumor size, advanced TNM stage, increased incidence of lymph node metastasis and shorter overall survival times in patients with NSCLC. Furthermore, CBR3­AS1­knockdown notably suppressed cellular proliferation, migration and invasiveness in vitro, and also promoted apoptosis and suppressed tumorigenicity in vivo. Mechanistic investigation demonstrated that CBR3­AS1 functions as a competing endogenous RNA for microRNA­509­3p (miR­509­3p) in NSCLC cells. Furthermore, miR­509­3p exerted tumor­suppressive effects in NSCLC, and histone deacetylase 9 (HDAC9) was identified as a direct target of miR­509­3p. HDAC9 expression was suppressed by CBR3­AS1 depletion, which was abolished by miR­509­3p inhibition. Further rescue experiments revealed that increasing the output of the miR­509­3p/HDAC9 axis counteracted the CBR3­AS1 depletion­induced inhibitory effects on NSCLC cells. Collectively, the results of the present study indicate that the CBR3­AS1/miR­509­3p/HDAC9 pathway exerts tumor­promoting actions in NSCLC oncogenesis and progression, suggesting that this pathway is an effective target for the management of NSCLC.

Inhibitory effect of trisodium citrate on biofilms formed by Klebsiella pneumoniae.

OBJECTIVES: Klebsiella pneumoniae is a significant nosocomial pathogen related to ventilator-associated pneumonia owing to biofilm formation. Trisodium citrate (TSC) has antibacterial activity, but there is little research on the effect of TSC on biofilm formed by K. pneumoniae. The aims of this study were to evaluate the inhibitory effect of 4% TSC on K. pneumoniae biofilm formation and to determine the best time of TSC addition for biofilm inhibition. METHODS: A total of 45 K. pneumoniae strains isolated from tracheal tip and sputum specimens were included. Modified Congo red agar was used to screen for biofilm production. Biofilm-positive strains were cultured for 4 days. TSC (4%) was added either initially or 3 days later. Crystal violet staining was used to quantify biofilm mass by measuring the optical density at 570 nm (OD570). Scanning electron microscopy (SEM) was used to observe biofilm morphology. RESULTS: The OD570 was significantly lower in the 4% TSC group than that in the no-TSC group during the 4-day experiment. Compared with addition of TSC after 3 days, initial TSC addition resulted in a significant absorbance decrease (Day 4, 0.63 ± 0.11 later-TSC group vs. 0.41 ± 0.16 initial-TSC group). As observed by SEM, bacteria were stacked most densely in the no-TSC group on Day 4. In contrast, few bacteria were observed when TSC was added initially, whilst bacteria were obviously dispersed when TSC was added after 3 days. CONCLUSION: TSC can inhibit K. pneumoniae biofilm formation and has the best effect when added initially.

The evaluation of adenotonsillectomy on TNF-α and IL-6 levels in obese children with obstructive sleep apnea.

OBJECTIVE: (1) To evaluate the effect of adenotonsillectomy (T&A) on clinical signs in obese children with obstructive sleep apnea (OSA); (2) to evaluate the changes in tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 levels pre- and post-surgery; and (3) to determine a potential explanation for changes in cytokine levels after T&A therapy. SUBJECTS AND METHODS: In brief, 90 obese children with OSA were enrolled. Polysomnographic data, such as an apnea-hypopnea index (AHI) and oxyhemoglobin saturation (SaO2), and demographic information, such as body mass index (BMI), were collected. TNF-α and IL-6 levels were determined before and 6 months after T&A treatment. RESULTS: It was observed that 61.1% of obese children with OSA were persistent after T&A therapy. In general, AHI (events/h) measures decreased from 22.26 ± 9.141 to 8.88 ± 5.909 before and after a 6-month follow-up period, respectively, and nadir SaO2 (%) levels increased from 74.26 ± 7.486 to 86.37 ± 5.578 pre- and post-surgery, respectively. However, no discernible differences in BMI measures before and after a 6-month follow-up period (40.996 ± 5.2486 vs. 40.597 ± 5.0540) were observed. No significant differences in the TNF-α and IL-6 levels pre- and post-surgery were observed. TNF-α and IL-6 plasma levels were highly correlated with BMI measurements (TNF-α: r(2)=0.2350, P<0.001; IL-6: r(2)=0.6629, P<0.001). In contrast, there was no association between plasma levels and AHI measures (TNF-α: r(2)=0.03377, P=0.0829; IL-6: r(2)=0.03946, P=0.0605). CONCLUSION: T&A therapy can improve clinical symptoms and signs but is not a cure for the treatment of obese children with OSA. The inflammation levels of TNF-α and IL-6 changed little. To reduce the risk for excessive daytime sleepiness, supplementary therapies should be introduced for OSA.

[Exploration of the correlation between Chinese medicine syndrome types of verrucous gastritis and the pressions of gastric mucosal hypoxia-inducible factor-1alpha as well as downstream molecules].

OBJECTIVE: To study the expressions changes of gastric mucosal hypoxia-inducible factor-1alpha (HIF-1alpha) and downstream molecules [such as vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2)] of verrucous gastritis (VG) patients of different Chinese medicine (CM) syndrome types and their clinical significance. METHODS: Totally 94 VG patients were assigned to Gan-Wei disharmony group (GWDG, 28 cases), the damp-heat in Pi-Wei group (DHPWG, 17 cases), the blood stasis in Wei-collateral group (BSWCG, 20 cases), and the insufficiency of Pi-yang group (IPYG, 29 cases). Another 30 patients with chronic mild non-active superficial gastritis patients accompanied with negative Hp infection were recruited as the control group. The Hp infection was detected using 14C-labeled urea breath test. The expressions of HIF-1alpha, VEGF, and COX-2 in the gastric mucosal tissue were detected using immunohistochemical EnVision two-step test. RESULTS: The positive Hp infection rate in VG patients was 37.23% (35/94 cases). The positive Hp infection rate in patients of DHPWG (76. 47%) was significantly higher than the other three groups (32.14% in GWDG, 31.03% in IPYG, and 20.00% in BSWCG, P < 0.01). The expressions of HIF-1alpha and COX-2 in VG patients of different syndrome types were obviously higher than those of the control group (P < 0.01, P < 0.05). Of them, the expression of HIF-1alpha was the highest in BSWCG and the expression of COX-2 was the highest in DHPWG. The expression of VEGF was higher in DHPWG and IPYG than in the control group and the GWDG (P < 0.01, P < 0.05). CONCLUSIONS: The expressions of HIF-1alpha, VEGF, COX-2, and Hp infection showed certain changes in VG patients of different syndrome types. The expression of HIF-1alpha was the strongest in BSWCG. The expressions of VEGF and COX-2 as well as Hp infection were the highest in DHPWG. All showed the specificity of CM syndromes.

Antibiotic resistance and molecular epidemiology of the beta-lactamase-producing Haemophilus influenzae isolated in Chongqing, China.

This study aimed to determine the antibiotic resistance and molecular epidemiology of Haemophilus influenzae isolated from children with acute respiratory infection in Chongqing, China. To this end, 1967 H. influenzae isolates from 2006 to 2009 were analysed regarding ß-lactamase production and antibiotic resistance. Ninety-nine ß-lactamase-producing H. influenzae isolates from 2010 were analysed for antibiotic resistance and promoter regions of bla(TEM) (-1) . ß-lactamase production was found in 35.8% (705/1967) of the strains. All ninety-nine ß-lactamase-producing strains from 2010 were of the TEM-1 type as determined by PCR but did not produce the predicted 1075 bp product. According to PCR-SSCP and DNA sequencing, the promoter regions of bla(TEM) (-1) were categorized into 6 genotypes as SSCP1 (Pdel), SSCP2 (Pa/Pb), SSCP3 (P4), SSCP4 (Prpt.b), SSCP5 (2Prpt) and SSCP6 (P3.b). The Pdel, Pa/Pb and Prpt.b were common promoters of bla(TEM) (-1) for H. influenzae isolated from children in Chongqing. Strains with Prpt.b were more resistant to ampicillin (AMP) than strains with Pdel, Pa/Pb and P4 (p < 0.05). Therefore, bla(TEM-1) ß-lactamase is the main mechanism for resistance of H. influenzae to ampicillin in Chongqing. Furthermore, the Prpt.b promoters may be related to the high resistance of H. influenzae to AMP.

Impact of adenotonsillectomy on high-sensitivity C-reactive protein levels in obese children with obstructive sleep apnea.

OBJECTIVE: To evaluate the effect of adenotonsillectomy (T&A) in obese children with obstructive sleep apnea (OSA) and to compare changes in high-sensitivity C-reactive protein (hs-CRP) levels before and 6 months after T&A in obese children with OSA. STUDY DESIGN: Before and after study with planned data collection. SETTING: Tertiary care, university-based pediatric hospital. SUBJECTS AND METHODS: Seventy-five obese children with OSA were included. Clinical information such as the apnea-hypopnea index (AHI), nadir oxyhemoglobin saturation (SaO(2)), and body mass index (BMI) were recorded. The hs-CRP level was determined before T&A and at the 6-month follow-up examination. RESULTS: Reductions in AHI (21.96 ± 9.277 before T&A vs 8.64 ± 5.997 after 6 months of T&A) and higher levels of nadir SaO(2) (74.08 ± 7.860 before T&A vs 86.87 ± 5.586 after 6 months of T&A) were observed. The hs-CRP levels were obviously correlated with BMI (r = 0.7948, P < .001). Other than AHI (r = 0.0579, P = .6217) in obese children, however, hs-CRP levels showed no changes 6 months after T&A therapy. CONCLUSION: T&A treatment improves clinical signs and symptoms in obese children but does not reduce chronic inflammation as reflected by hs-CRP. To lower the risks of cardiovascular disease and diabetes mellitus morbidity, other treatments should be taken into account.