An autophagy-inducing stapled peptide induces mitochondria dysfunction and triggers autotic cell death in triple-negative breast cancer.

Autophagy is a lysosome-dependent bulk degradation process essential for cell viability but excessive autophagy leads to a unique form of cell death termed autosis. Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer with notable defect in its autophagy process. In previous studies, we developed stapled peptides that specifically targeted the essential autophagy protein Beclin 1 to induce autophagy and promote endolysosomal trafficking. Here we show that one lead peptide Tat-SP4 induced mild increase of autophagy in TNBC cells but showed potent anti-proliferative effect that could not be rescued by inhibitors of programmed cell death pathways. The cell death induced by Tat-SP4 showed typical features of autosis including sustained adherence to the substrate surface, rupture of plasma membrane and effective rescue by digoxin, a cardioglycoside that blocks the Na+/K+ ATPase. Tat-SP4 also induced prominent mitochondria dysfunction including loss of mitochondria membrane potential, elevated mitochondria reactive oxygen species and reduced oxidative phosphorylation. The anti-proliferative effect of Tat-SP4 was confirmed in a TNBC xenograft model. Our study uncovers three notable aspects of autosis. Firstly, autosis can be triggered by moderate increase in autophagy if such increase exceeds the endogenous capacity of the host cells. Secondly, mitochondria may play an essential role in autosis with dysregulated autophagy leading to mitochondria dysfunction to trigger autosis. Lastly, intrinsic autophagy deficiency and quiescent mitochondria bioenergetic profile likely render TNBC cells particularly susceptible to autosis. Our designed peptides like Tat-SP4 may serve as potential therapeutic candidates against TNBC by targeting this vulnerability.

Resveratrol improves the cytotoxic effect of CD8 +T cells in the tumor microenvironment by regulating HMMR/Ferroptosis in lung squamous cell carcinoma.

Ferroptosis, an iron-dependent cell death process, is a potential therapeutic strategy for Lung squamous cell carcinoma (LUSC). Resveratrol (RES) is an anti-tumor polyphenol. However, whether and how RES treats LUSC is not yet known. This study aimed to investigate the effect of RES on LUSC and to explore its potential mechanism. This study used a combination of proteomics, bioinformatics, clinical samples, and cell experiments to study the interaction between HMMR and the ferroptosis signaling pathway and investigate the role of RES in regulating tumor immune microenvironment and anti-tumor by cytotoxic CD8 +T cells in LUSC. Ferroptosis signaling pathway and HMMR were involved in the LUSC tumor immune microenvironment and correlated with worse prognosis of LUSC patients. RES+H520 cells induced a higher level of ferroptosis and MDA, mainly by reducing the expression of GPX4 and SLC7A11, inducing the expression of ACSL4 and TFRC. HMMR, GSH, and SOD contents were lower observed than in H520 cells. When HMMR was expressed, SLC7A11 was also highly expressed in LUSC, and there was an interaction between HMMR expression and SLC7A11. In addition, RES increased the TNF-α, IFN-γ, IL-12, and IL-2 expression and increased the cytotoxic effects of CD8 +T cells expressions in LUSC. Resveratrol regulates SLC7A11-HMMR interaction, activates ferroptosis, enhances the cytotoxic effect of CD8 +T cells, and regulates the tumor immune microenvironment. Based on the pathogenesis of LUSC and the clinical efficacy of RES, this study explored the influence of RES on LUSC, clarified its biological effects, and further provided cell biological basis for the clinical application of RES, which could guide clinical combination and personalized medicine, improve the response rate of immunotherapy and benefit more patients with LUSC.

Identification and validation of Osteopontin and receptor for hyaluronic acid-mediated motility (RHAMM, CD168) for potential immunotherapeutic significance of in lung squamous cell carcinoma.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths. Immunotherapy is a promising therapeutic approach, but the population best suited to immunotherapy is yet to be determined. MATERIALS AND METHODS: Lung squamous cell carcinoma (LUSC) was chosen as the object for the present study. Four gene expression profiles were retrieved from the GEO database. 141 differentially expressed genes (DEGs) were detected in LUSC tissues and normal tissues by the GEO2R tool and Venn diagram software. RESULTS: 34 candidate genes were selected for further analysis. A Kaplan-Meier survival plot further isolated 29 of 34 genes and after re-validation using gene expression profiling interactive analysis and pathway enrichment, Bonferroni correction was used to adjust P values, results showed that two genes (CD168 and OPN) were markedly enriched in the extracellular matrix (ECM)-receptor interaction pathway. We believe this pathway and genes may be tightly involved in the LUSC tumor immune microenvironment. We conducted a further cellular study to knock-down OPN in H520 cells using siRNA. The expression of CD168 was reduced in siRNA-OPN H520 cells (P < 0.05). Our results indicate that the arrest of CD168 occurs after the downregulation of the OPN protein, suggesting that OPN participates in ECM-receptor interactions. CONCLUSIONS: By using integrated bioinformatics, we have identified CD168 and OPN as DEGs with poor prognosis in LUSC and have validated their interaction in the ECM receptor pathway. These genes could be potential diagnostic and therapeutic targets for LUSC patients undergoing immunotherapy.

The Ferroptosis-NLRP1 Inflammasome: The Vicious Cycle of an Adverse Pregnancy.

One of the hallmarks of placental dysfunction is the increase of oxidative stress. This process, along with the overexpression of the inflammasome, creates a downward spiral that can lead to a series of severe pregnancy complications. Ferroptosis is a form of iron-mediated cell death involving the accumulation of reactive oxygen species, lipid peroxides. In this study, the rats' model of oxidative stress abortion was established, and hydrogen peroxide (H2O2) was used to establish a cellular model of placental oxidative stress. RNAi, western blot, and immunofluorescence were used to evaluate the expression of specific markers of ferroptosis and the expression of the inflammasome in placental trophoblast cells. We observed excessive levels of ferroptosis and inflammasome activation in both rats' model and placental trophoblast cell model of oxidative stress. When the NLRP1 inflammasome was silenced, the expression levels of GSH and Glutathione peroxidase 4 (GPX4) were increased, while the expression levels of transferrin receptor 1 (TFR1), acyl-CoA synthetase long-chain family member 4 (ACSL4), Superoxide dismutase (SOD), and Malondialdehyde (MDA) were decreased. However, when an NLRP1 activator was applied, we observed the opposite phenomenon. We further explored the mechanisms underlying the actions of ferroptosis to inflammasomes. The expression levels of NLRP1, NLRP3, IL-1ß, and caspase-1 were positively correlated with the ferroptosis following the application of ferroptosis inhibitor (ferrostatin-1) and ferroptosis activator (erastin). The existence of ferroptosis was demonstrated in the oxidative stress model of placental trophoblast cells; the results also indicate ferroptosis is linked with the expression of NLRP1 inflammasome. These findings may provide a valuable therapeutic target for the pathogenesis of pregnancy-related diseases.

A possible interaction between linezolid and digoxin: A case report of therapeutic drug monitoring.

Drug-drug interactions lead to altered clinical effects, including adverse reactions. Therapeutic drug monitoring of digoxin is necessary due to its narrow therapeutic range. Linezolid can cause variable exposures in patients hospitalized in the intensive care unit owing to its possibility of drug-drug interactions. We present a patient with pneumonia and heart failure who experienced a possible drug interaction between linezolid and digoxin, resulting in high serum concentrations of both drugs. Also, the patient developed thrombocytopenia likely related to linezolid. The linezolid dose required to maintain sufficient levels had to reduce to 50% of the usual linezolid dose. A quarter dose of the standard digoxin dose was needed. Although the underlying mechanism of the drug interaction is unclear, we recommend conducting therapeutic drug monitoring when linezolid and digoxin are administered concurrently.

COVID-19 Infections and Deaths among Connecticut Nursing Home Residents: Facility Correlates.

BACKGROUND/OBJECTIVES: To determine the associations of nursing home registered nurse (RN) staffing, overall quality of care, and concentration of Medicaid or racial and ethnic minority residents with 2019 coronavirus disease (COVID-19) confirmed cases and deaths by April 16, 2020, among Connecticut nursing home residents. DESIGN: Cross-sectional analysis on Connecticut nursing home (n = 215) COVID-19 report, linked to other nursing home files and county counts of confirmed cases and deaths. Multivariable two-part models determined the associations of key nursing home characteristics with the likelihood of at least one confirmed case (or death) in the facility, and with the count of cases (deaths) among facilities with at least one confirmed case (death). SETTING: All Connecticut nursing homes (n = 215). PARTICIPANTS: None. INTERVENTION: None. MEASUREMENTS: Numbers of COVID-19 confirmed cases and deaths among residents. RESULTS: The average number of confirmed cases was eight per nursing home (zero in 107 facilities), and the average number of confirmed deaths was 1.7 per nursing home (zero in 131 facilities). Among facilities with at least one confirmed case, every 20-minute increase in RN staffing (per resident day) was associated with 22% fewer confirmed cases (incidence rate ratio [IRR] = .78; 95% confidence interval [CI] = .68-.89; P < .001); compared with one- to three-star facilities, four- or five-star facilities had 13% fewer confirmed cases (IRR = .87; 95% CI = .78-.97; P < .015), and facilities with high concentration of Medicaid residents (IRR = 1.16; 95% CI = 1.02-1.32; P = .025) or racial/ethnic minority residents (IRR = 1.15; 95% CI = 1.03-1.29; P = .026) had 16% and 15% more confirmed cases, respectively, than their counterparts. Among facilities with at least one death, every 20-minute increase in RN staffing significantly predicted 26% fewer COVID-19 deaths (IRR = .74; 95% CI = I .55-1.00; P = .047). Other focused characteristics did not show statistically significant associations with deaths. CONCLUSION: Nursing homes with higher RN staffing and quality ratings have the potential to better control the spread of the novel coronavirus and reduce deaths. Nursing homes caring predominantly for Medicaid or racial and ethnic minority residents tend to have more confirmed cases.

Effect of combined spinal-epidural analgesia in labor on frequency of emergency cesarean delivery among nulliparous Chinese women.

OBJECTIVE: To determine whether combined spinal-epidural analgesia (CSEA) during labor increases the frequency of emergency cesarean delivery among Chinese nulliparous women. METHODS: In a retrospective study, the medical records of nulliparous women with a singleton fetus in cephalic presentation who delivered at term at Tongling Maternity Hospital, China, between January 2012 and December 2014 were reviewed. Information about CSEA, mode of delivery, labor duration, oxytocin augmentation, and neonatal outcome was obtained. Logistic regression was used to examine independent associations between CSEA and emergency cesarean after controlling for confounding variables. RESULTS: Among 3456 women included, 1786 (51.7%) received CSEA and 1670 (48.3%) received no labor analgesia. Emergency cesarean was more frequent among CSEA users (219/1786 [12.3%]) than non-users (119/1670 [7.1%]; P<0.001). Among the maternal-fetal variables included in multivariate regression, maternal age, maternal height, cervical dilatation at admission, birth weight, and CSEA use were significantly associated with emergency cesarean. After adjustment, women with CSEA maintained a slightly increased risk for cesarean (adjusted odds ratio 1.54, 95% confidence interval 1.20-2.00). CONCLUSION: Among Chinese nulliparous women, use of CSEA for labor pain was associated with an increased risk of emergency cesarean delivery; moreover, this effect was maintained after adjustment for other potential obstetric risk factors.

[Research on Test Method of Metallic Element Contained in Tea Based on EDXRF Technique].

As it has been certified by experimental testing that when using the energy-dispersive X-ray fluorescence (EDXRF) method to analyze the metallic elements contained in the tea the energy segment of effective X-ray fluorescence photons is located between 3 and 16 keV. Accordingly the spectral correction element is targeted at the copper elements located near the energy center(8 keV). The copper elements are also used as the picketage to be the standard curve. In the energy segment of effective X-ray fluorescence photons contained in the tea 1.25 mg · kg(-1) of the average detection limit was obtained by using the spiked method to analyze four elements of copper, iron, zinc and lead. Compared with the flame atomic absorption spectrum(FAAS), the actual relative error of the tested value by EDXRF is less than 6%, and the relative standard deviation is less than 5%. The result by T test shows that p > 0.05. The conclusions are that there are no statistically significant differences between EDXRF and FAAS. The measured results gained by the two methods agree with each other. And EDXRF can be used thoroughly to test the metal contents contained in the tea. The result shows that it is feasible to test the metallic contents contained in the tea by EDXRF, and its measured result can meet the requirements of field testing and analysis.

Effects of miR-9 and tetramethylpyrazine on activation of hepatic stellate cells.

Micro-RNAs (miRNAs) are involved in regulation of the incidence and development of several hepatic diseases. Thus manipulating miRNAs may be a promising therapeutic strategy against these entities. In this study hepatic stellate cells (HSCs) were transfected with hsa-miR-9 or anti-hsa-miR-9, treated with tetramethylpyrazine (TMP), or subjected to treatment with TMP and hsa-miR-9 transfection (combined treatment group). Then, real-time polymerase chain reaction (PCR) was performed to measure mRNA levels of hsa-miR-9. Expression of hsa-miR-9 was highest in the combination treatment group compared with other groups, and significantly higher than TMP-treated and hsa-miR-9-transfected groups (both p<0.05). The anti-hsa-miR-9-transfected group expressed the lowest mRNA level of hsa-miR-9 with marked decrease versus control (p<0.05). Downstream factors that may be affected by miR-9 such as leptin, α-smooth muscle actin (SMA), and collagen I, as well as phosphorylation levels of Janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3) were investigated at the protein level. All these factors were regulated contrariwise to expression trends of hsa-miR-9, showing the lowest level in the combination treatment group and highest level in anti-hsa-miR-9-transfected group. These results suggest that both transfection of hsa-miR-9 and TMP can lead to upregulated endogenous expression of hsa-miR-9, inhibit activation of JAK1/STAT3 signal pathway induced by leptin, and lead to reduction of α-SMA and collagen I-thus impeding activation of HSC.

[Genome-scale sequence data processing and epigenetic analysis of DNA methylation].

A new approach recently developed for detecting cytosine DNA methylation (mC) and analyzing the genome-scale DNA methylation profiling, is called BS-Seq which is based on bisulfite conversion of genomic DNA combined with next-generation sequencing. The method can not only provide an insight into the difference of genome-scale DNA methylation among different organisms, but also reveal the conservation of DNA methylation in all contexts and nucleotide preference for different genomic regions, including genes, exons, and repetitive DNA sequences. It will be helpful to under-stand the epigenetic impacts of cytosine DNA methylation on the regulation of gene expression and maintaining silence of repetitive sequences, such as transposable elements. In this paper, we introduce the preprocessing steps of DNA methylation data, by which cytosine (C) and guanine (G) in the reference sequence are transferred to thymine (T) and adenine (A), and cytosine in reads is transferred to thymine, respectively. We also comprehensively review the main content of the DNA methylation analysis on the genomic scale: (1) the cytosine methylation under the context of different sequences; (2) the distribution of genomic methylcytosine; (3) DNA methylation context and the preference for the nucleotides; (4) DNA- protein interaction sites of DNA methylation; (5) degree of methylation of cytosine in the different structural elements of genes. DNA methylation analysis technique provides a powerful tool for the epigenome study in human and other species, and genes and environment interaction, and founds the theoretical basis for further development of disease diagnostics and therapeutics in human.

[Tissue culture of medicinal plant and abscisic acid].

Abscisic acid (ABA) plays a key role in many physiological processes of plants, and it was also applied to fields of medicinal plant biotechnology. The article presents a review of some recent application of ABA in enhancing the production of secondary metabolites of medicinal plants, improving the in vitro conservation in medicinal plant tissue culture system.

Neurological manifestations of Takayasu arteritis.

OBJECTIVE: To investigate the clinical neurological manifestations of Takayasu arteritis (TA). METHODS: A retrospective study was conducted with 63 consecutive TA cases admitted to Peking Union Medical College Hospital from January 2009 to May 2010. All the patients fulfilled the diagnostic criteria of TA by the American College of Rheumatology. Among the 63 TA patients, 27 with neurological manifestations were included in the present study. All the patients were evaluated using standardized neurological examination, sonography, computed tomography (CT) angiography, and cerebral CT or magnetic resonance imaging. RESULTS: Dizziness and visual disturbance were the most common symptoms, which occurred in 20 (74.1%) and 16 (59.3%) patients respectively. Another common symptom was headache, observed in 15 (55.6%) patients. Six (22.2%) patients had suffered from ischemic stroke; 7 (25.9%) patients had epileptic seizures. Two (7.4%) patients were diagnosed as reversible posterior encephalopathy syndrome (RPES) based on typical clinical and imaging manifestations. CONCLUSIONS: Neurological manifestations are common symptoms in TA patients in the chronic phase, including dizziness, visual disturbance, headache, ischemic stroke, seizures, and some unusual ones such as RPES. We suggested RPES be included into the differential diagnosis of acute neurological changes in TA.

Angiotensin-(1-7) attenuates hyposmolarity-induced ANP secretion via the Na+-K+ pump.

The alteration in osmolarity challenges cell volume regulation, a vital element for cell survival. Hyposmolarity causes an increase in cell volume. Recently, it has been reported that the renin-angiotensin system (RAS) plays a role in cell volume regulation. We investigated the effect of angiotensin-(1-7) [Ang-(1-7)] on hyposmolarity-induced atrial natriuretic peptide (ANP) secretion in normal and diabetic (DM) rat atria and modulation of the effect of Ang-(1-7) by the Na(+)-K(+) pump. Using isolated control rat atria, we observed that perfusion of hyposmotic solution into the atria increased ANP secretion. When Ang-(1-7) [0.1 microM or 1 microM] was perfused in a hyposmolar solution, it decreased the hyposmolarity-induced ANP secretion in a dose-dependent manner. This effect of Ang-(1-7) could be mediated by the Na(+)-K(+) pump, since ouabain, an Na(+)-K(+) pump inhibitor, significantly decreased the effect of Ang-(1-7) on hyposmolarity-induced ANP secretion. In contrast, N(omega) Nitro-l-arginine methyl ester hydrochloride (l-NAME) did not modify the effect of Ang-(1-7) on the hyposmolarity-induced ANP secretion. Interestingly, the ANP secretion was increased robustly by the perfusion of the hyposmolar solution in the DM atria, as compared to the control atria. However, the inhibitory effect of Ang-(1-7) on the hyposmolarity-induced ANP secretion was not observed in the DM atria. In the DM atria, atrial contractility was significantly increased. Taken together, we concluded that Ang-(1-7) attenuated hyposmolarity-induced ANP secretion via the Na(+)-K(+) pump and a lack of Ang-(1-7) response in DM atria may partly relate to change in Na(+)-K(+) pump activity.

De novo assembly of human genomes with massively parallel short read sequencing.

Next-generation massively parallel DNA sequencing technologies provide ultrahigh throughput at a substantially lower unit data cost; however, the data are very short read length sequences, making de novo assembly extremely challenging. Here, we describe a novel method for de novo assembly of large genomes from short read sequences. We successfully assembled both the Asian and African human genome sequences, achieving an N50 contig size of 7.4 and 5.9 kilobases (kb) and scaffold of 446.3 and 61.9 kb, respectively. The development of this de novo short read assembly method creates new opportunities for building reference sequences and carrying out accurate analyses of unexplored genomes in a cost-effective way.

Counting of oligomers in sequences generated by markov chains for DNA motif discovery.

By means of the technique of the imbedded Markov chain, an efficient algorithm is proposed to exactly calculate first, second moments of word counts and the probability for a word to occur at least once in random texts generated by a Markov chain. A generating function is introduced directly from the imbedded Markov chain to derive asymptotic approximations for the problem. Two Z-scores, one based on the number of sequences with hits and the other on the total number of word hits in a set of sequences, are examined for discovery of motifs on a set of promoter sequences extracted from A. thaliana genome. Source code is available at http://www.itp.ac.cn/zheng/oligo.c.

1-Phenyl-2-trifluoro-methyl-4-quinolone.

In the title mol-ecule, C(16)H(10)F(3)NO, the N-bound phenyl ring is oriented nearly orthogonal to the quinolinyl ring in order to avoid steric clashes with the trifluoro-methyl substituent [dihedral angle 89.7 (1)°].