DNA Extraction and Comparison between Old and Fresh Necrophilic Fly Samples.

A total of five samples of Chrysomya megacephala samples - three fresh samples, one sample stored in alcohol for 2 years, and one sample stored in dry sealed storage for 2 years protected from light only - were selected to investigate whether a blood DNA extraction kit could extract DNA from necrophilous flies and to determine whether alcohol could prolong the preservation of necrophilous flies' DNA. First, the blood DNA extraction kit was used to extract DNA from their thorax tissues. Then, the DNA purity and concentration were examined using a microplate reader and a fluorometer. Finally, PCR amplification and electrophoresis of the extracted DNA were done with necrophilic fly-specific primers located in the mitochondrial CO I gene sequence. The results showed that the DNA purity of all samples was greater than 2.0. The DNA concentration was observed to be of the following order: fresh samples > alcohol-preserved old samples > untreated, old samples. All samples had specific electrophoretic bands after PCR amplification. In conclusion, a blood DNA extraction kit can be used to extract DNA from necrophilic flies successfully, and the DNA concentration of fresh fly samples is greater than that of old fly samples. The flies can be stored in alcohol for a long time.

The Impacts of Planting Patterns Combined with Irrigation Management Practices on Watermelon Growth, Photosynthesis, and Yield.

(1) Background: Crop yields in China's arid and semi-arid regions are limited by water shortages. Exploring the interactions and resource utilization among agroforestry species is key to maintaining diversified agricultural production. (2) Objective: An apple-watermelon agroforestry system and watermelon sole-cropping system were compared to quantify how resource availability (light, water) and watermelon performance (leaf photosynthetic rate, growth, and yield) change with irrigation strategies. (3) Methods: A three-year apple and watermelon field experiment was conducted in a young apple orchard in the arid area of central Ningxia to test the effect of light competition and irrigation systems on light environment, leaf photosynthetic rate, plant growth, and yield in watermelon. The experiment encompassed two planting patterns: (i) apple-watermelon agroforestry (AF) and watermelon sole-cropping (SC) and (ii) three irrigation quotas (W1: 105 mm, W2: 210 mm, and W3: 315 mm). (4) Results: The results show that the agroforestry planting pattern extended the growth period of watermelon and increased the leaf area index. Mean daily shade intensity increased by 16.02% from 2020 to 2022. The land equivalent ratio (LER) was >1 in 2021 and 2022. The SWC, leaf photosynthetic rate, LAI, and yield of watermelon in an agroforestry planting pattern were lower than when in a sole-cropping planting pattern. However, under the W1 irrigation strategy, the total soluble solids of the agroforestry planting pattern were 2.27% higher than those of the sole-cropping pattern, and the yield of the agroforestry planting pattern was 2.59% higher than that of the sole-cropping pattern. Under the W3 irrigation strategy, the average watermelon weight in the agroforestry planting pattern was 2.85% higher than that of the sole-cropping pattern. A path analysis showed that the agroforestry planting pattern can increase the yield by increasing soil water content, which is different from the sole-cropping pattern. (5) Conclusions: The results confirm that the apple-watermelon agroforestry planting pattern reduced watermelon yields. However, the LER of the agroforestry system was greater than 1.0. It is reasonable to plant watermelons in young apple forests.

Association between the triglyceride glucose index and length of hospital stay in patients with heart failure and type 2 diabetes in the intensive care unit: a retrospective cohort study.

Background: The coexistence of heart failure and diabetes is prevalent, particularly in Intensive Care Units (ICU). However, the relationship between the triglyceride-glucose (TyG) index, heart failure, diabetes, and the length of hospital stay (LHS) in patients with cerebrovascular disease in the ICU remains uncertain. This study aims to investigate the association between the TyG index and LHS in patients with heart failure and diabetes. Methods: This retrospective study utilized the Medical Information Mart for Intensive Care (MIMIC)-IV database to analyze patients with diabetes and heart failure. Participants were categorized into quartiles based on the TyG index, and the primary outcome was LHS. The association between the TyG index at ICU admission and LHS was examined through multivariable logistic regression models, restricted cubic spline regression, and subgroup analysis. Results: The study included 635 patients with concurrent diabetes and heart failure. The fully adjusted model demonstrated a positive association between the TyG index and LHS. As a tertile variable (Q2 and Q3 vs Q1), the beta (ß) values were 0.88 and 2.04, with a 95% confidence interval (95%CI) of -0.68 to 2.44 and 0.33 to 3.74, respectively. As a continuous variable, per 1 unit increment, the ß (95% CI) was 1.13 (0.18 to 2.08). The TyG index's relationship with LHS showed linearity (non-linear p = 0.751). Stratified analyses further confirmed the robustness of this correlation. Conclusion: The TyG index exhibited a linearly positive association with the LHS in patients with both heart failure and diabetes. Nevertheless, prospective, randomized, controlled studies are imperative to substantiate and validate the findings presented in this investigation.

A p21-ATD mouse model for monitoring and eliminating senescent cells and its application in liver regeneration post injury.

Cellular senescence associates with pathological aging and tissue dysfunctions. Studies utilizing mouse models for cell lineage tracings have emphasized the importance of senescence heterogeneity in different organs and cell types. Here, we constructed a p21- (Akaluc - tdTomato - Diphtheria Toxin Receptor [DTR]) (ATD) mouse model to specifically study the undefined mechanism for p21-expressing senescent cells in the aged and liver injury animals. The successful expressions of these genes enabled in vitro flow cytometric sorting, in vivo tracing, and elimination of p21-expressing senescent cells. During the natural aging process, p21-expressing cells were found in various tissues of p21-ATD mice. Eliminating p21-expressing cells in the aged p21-ATD mice recovered their multiple biological functions. p21-ATD/Fah-/- mice, bred from p21-ATD mice and fumarylacetoacetate hydrolase (Fah)-/- mice of liver injury, showed that the majority of their senescent hepatocytes were the phenotype of p21+ rather than p16+. Furthermore, eliminating the p21-expressing hepatocytes significantly promoted the engraftment of grafted hepatocytes and facilitated liver repopulation, resulting in significant recovery from liver injury. Our p21-ATD mouse model serves as an optimal model for studying the pattern and function of p21-expressing senescent cells under the physical and pathological conditions during aging.

Uncommon Presentation of Recurrent Lung Adenocarcinoma: A Finger Ulcer Induced by Subclavian Artery Invasion Successfully Healed With Viabahn VBX Treatment.

Recurrence of a lung tumor invading the subclavian artery, causing stenosis and leading to finger ulcers as the initial symptom, is rare. We employed endovascular techniques, inserting a Viabahn® VBX covered stent (W. L. Gore & Associates, Flagstaff, Arizona) to aid in ulcer healing and improve the patient's quality of life. The patient, a 73-year-old male, had a history of lung adenocarcinoma resection two years prior but had not undergone follow-up examinations or cancer-specific treatments. Clinical examination revealed an invasion of the right subclavian artery by the recurrent tumor, resulting in severe stenosis and ischemic symptoms in the right upper limb. Given the patient's advanced cancer stage and the decline of further tumor-specific treatments, an endovascular intervention using a Viabahn VBX covered stent was performed to improve blood flow and promote ulcer healing. The stent demonstrated exceptional stability and patency during the six-month follow-up, greatly improving the patient's quality of life. This case highlights the importance of recognizing atypical symptoms as potential indicators of tumor recurrence or progression and demonstrates the promising role of covered stents in managing vascular complications in selected patients with advanced-stage malignancies.

Bacillus velezensis SQR9 promotes plant growth through colonization and rhizosphere-phyllosphere bacteria interaction.

The rhizosphere and phyllosphere of plants are home to a diverse range of microorganisms that play pivotal roles in ecosystem services. Consequently, plant growth-promoting bacteria (PGPB) are extensively utilized as inoculants to enhance plant growth and boost productivity. Despite this, the interactions between the rhizosphere and phyllosphere, which are influenced by PGPB inoculation, have not been thoroughly studied to date. In this study, we inoculated Bacillus velezensis SQR9, a PGPB, into the bulk soil, rhizosphere or phyllosphere, and subsequently examined the bacterial communities in the rhizosphere and phyllosphere using amplicon sequencing. Our results revealed that PGPB inoculation increased its abundance in the corresponding compartment, and all treatments demonstrated plant growth promotion effects. Further analysis of the sequencing data indicated that the presence of PGPB exerted a more significant impact on bacterial communities in both the rhizosphere and phyllosphere than in the inoculation compartment. Notably, the PGPB stimulated similar rhizosphere-beneficial microbes regardless of the inoculation site. We, therefore, conclude that PGPB can promote plant growth both directly and indirectly through the interaction between the rhizosphere and phyllosphere, leading to the enrichment of beneficial microorganisms.

Biological factors driving colorectal cancer metastasis.

Approximately 20% of colorectal cancer (CRC) patients present with metastasis at diagnosis. Among Stage I-III CRC patients who undergo surgical resection, 18% typically suffer from distal metastasis within the first three years following initial treatment. The median survival duration after the diagnosis of metastatic CRC (mCRC) is only 9 mo. mCRC is traditionally considered to be an advanced stage malignancy or is thought to be caused by incomplete resection of tumor tissue, allowing cancer cells to spread from primary to distant organs; however, increasing evidence suggests that the mCRC process can begin early in tumor development. CRC patients present with high heterogeneity and diverse cancer phenotypes that are classified on the basis of molecular and morphological alterations. Different genomic and nongenomic events can induce subclone diversity, which leads to cancer and metastasis. Throughout the course of mCRC, metastatic cascades are associated with invasive cancer cell migration through the circulatory system, extravasation, distal seeding, dormancy, and reactivation, with each step requiring specific molecular functions. However, cancer cells presenting neoantigens can be recognized and eliminated by the immune system. In this review, we explain the biological factors that drive CRC metastasis, namely, genomic instability, epigenetic instability, the metastatic cascade, the cancer-immunity cycle, and external lifestyle factors. Despite remarkable progress in CRC research, the role of molecular classification in therapeutic intervention remains unclear. This review shows the driving factors of mCRC which may help in identifying potential candidate biomarkers that can improve the diagnosis and early detection of mCRC cases.

Revisiting the relationships between energy consumption, economic development and urban size: A global perspective using remote sensing data.

Existing methods of measuring energy consumption require complex statistics and computing. A real-time and globally applicable approach for comparing energy consumption across different cities is still lacking. Additionally, the nonlinear relationships and varying thresholds of energy consumption in relation to economic activities and urbanization remain unconfirmed. This study aims to fill these gaps by utilizing Suomi National Polar-orbiting Partnership Visible Infrared Imaging Radiometer Suite (NPP-VIIRS) nighttime light data in 2015 and a top-down approach based on a multiple regression model to examine energy consumption in global cities employing a redefined urban boundary. It also explores the accurate relationship between energy consumption, population density (as a proxy of urbanization), and per capita gross domestic product (GDP) across different regions and urban sizes using generalized additive models and regression models. High-resolution gridded population and GDP datasets covering the entire planet are utilized for this purpose. The study also estimates the development potentiality. The study yields followings outcomes: Firstly, the top 30 cities with the highest per capita energy consumption account for over 0.66% of the total per capita energy consumption of all cities. Secondly, in East Asia (EA) and Southeast Asia (SEA), the per capita energy consumption decreases when per capita GDP reaches $40,000 and $75,000, respectively, while it remains stable in cities located in Western Europe (WE) and North America (NA) as per capita GDP increases. Thirdly, the per capita energy consumption declines with increasing urban population density until reaching 10,000 person/km2, 22,000 person/km2, and 4000 person/km2 in EA, SEA, and NA, respectively. Fourthly, in Central Asia (CA), megacities can save over 100 Mbtu/population when per capita GDP increases by $1000 compared to big cities. This pioneering study provides a comparable investigation of energy consumption at the global city level, exploring its relationship with urbanization and economy by employing a unified calculation standard. It will facilitate long-term energy-saving policies and urban planning strategies.

Human respiratory syncytial virus subgroups A and B outbreak in a kindergarten in Zhejiang Province, China, 2023.

Background: In May-June 2023, an unprecedented outbreak of human respiratory syncytial virus (HRSV) infections occurred in a kindergarten, Zhejiang Province, China. National, provincial, and local public health officials investigated the cause of the outbreak and instituted actions to control its spread. Methods: We interviewed patients with the respiratory symptoms by questionnaire. Respiratory samples were screened for six respiratory pathogens by real-time quantitative polymerase chain reaction (RT-PCR). The confirmed cases were further sequenced of G gene to confirm the HRSV genotype. A phylogenetic tree was reconstructed by maximum likelihood method. Results: Of the 103 children in the kindergarten, 45 were classified as suspected cases, and 25 cases were confirmed by RT-PCR. All confirmed cases were identified from half of classes. 36% (9/25) were admitted to hospital, none died. The attack rate was 53.19%. The median ages of suspected and confirmed cases were 32.7 months and 35.8 months, respectively. Nine of 27 confirmed cases lived in one community. Only two-family clusters among 88 household contacts were HRSV positive. A total of 18 of the G gene were obtained from the confirmed cases. Phylogenetic analyses revealed that 16 of the sequences belonged to the HRSV B/BA9 genotype, and the other 2 sequences belonged to the HRSV A/ON1 genotype. The school were closed on June 9 and the outbreak ended on June 15. Conclusion: These findings suggest the need for an increased awareness of HRSV coinfections outbreak in the kindergarten, when HRSV resurges in the community after COVID-19 pandemic.

Cumulative live birth rate of in vitro fertilization cycle via progestin-primed ovarian stimulation versus gonadotropin-releasing hormone antagonist protocol in infertile women with normal ovarian reserve: an open-label, randomized controlled trial.

This study aimed to evaluate the cumulative live birth rate (cLBR) of progestin-primed ovarian stimulation (PPOS) protocol versus gonadotropin-releasing hormone antagonist (GnRH-ant) protocol for in vitro fertilization (IVF) cycle in infertile women with normal ovarian reserve (NOR). Infertile women with NOR who underwent their first IVF cycle were enrolled in an open-label randomized controlled trial. Patients were randomly assigned 1:1 to receive a freeze-all strategy with delayed embryo transfer (PPOS group, n = 174) and fresh embryo transfer first (GnRH-ant group, n = 174). The primary outcome was the cLBR per aspiration. The cLBR between the PPOS group and GnRH-ant group were comparable (55.75% vs. 52.87%, p = 0.591). A premature luteinizing hormone surge was not observed in the PPOS group, while there were six cases (3.45%) in the GnRH-ant group, but no premature ovulation in either of the groups. The pregnancy outcomes, including implantation rate, clinical pregnancy rate and miscarriage rate, were all comparable. In addition, the number of retrieved oocytes, mature oocytes and viable embryos were similar (all p > 0.05) between the two groups.

Observation and quantification of the pseudogap in unitary Fermi gases.

The microscopic origin of high-temperature superconductivity in cuprates remains unknown. It is widely believed that substantial progress could be achieved by better understanding of the pseudogap phase, a normal non-superconducting state of cuprates1,2. In particular, a central issue is whether the pseudogap could originate from strong pairing fluctuations3. Unitary Fermi gases4,5, in which the pseudogap-if it exists-necessarily arises from many-body pairing, offer ideal quantum simulators to address this question. Here we report the observation of a pair-fluctuation-driven pseudogap in homogeneous unitary Fermi gases of lithium-6 atoms, by precisely measuring the fermion spectral function through momentum-resolved microwave spectroscopy and without spurious effects from final-state interactions. The temperature dependence of the pairing gap, inverse pair lifetime and single-particle scattering rate are quantitatively determined by analysing the spectra. We find a large pseudogap above the superfluid transition temperature. The inverse pair lifetime exhibits a thermally activated exponential behaviour, uncovering the microscopic virtual pair breaking and recombination mechanism. The obtained large, temperature-independent single-particle scattering rate is comparable with that set by the Planckian limit6. Our findings quantitatively characterize the pseudogap in strongly interacting Fermi gases and they lend support for the role of preformed pairing as a precursor to superfluidity.

Vortex-assisted hydrophobic natural deep eutectic solvent liquid-liquid microextraction for the removal of silver ions from environmental water.

This study presents a novel approach for the quantification of silver ions in environmental water through the utilization of liquid-liquid microextraction, employing natural deep eutectic solvents in conjunction with inductively coupled plasma emission spectroscopy. The extracted solvent was characterized by Fourier transform infrared spectroscopy (FT-IR). The impact of various extractant types, extractant molar ratio, extractant volume, extraction time, and salt concentration on the efficacy of silver ion extraction was investigated. The findings indicate that the optimal extraction efficiency was attained by utilizing a 5-mL aqueous solution volume, containing 1000 µL thymol/lactic acid NADES 1:3, a salt concentration of 1 mg mL-1, a pH value of 4, and a vortex time of 4 min. Upon implementing the optimized experimental conditions, the recovery of target metal ions was from 96.9 to 101.0%. The relative standard deviations were observed to be within the range of 1.5 to 2.7%. The present study demonstrates the reproducibility, accuracy, and reliability of the method for detecting silver ions in environmental water, with linear range of 5~1000 ng mL-1 and limits of detection (LOD) and limits of quantification (LOQ) of 1.52 ng mL-1 and 5.02 ng mL-1, respectively.

Long-Term Outcomes in Older Patients with Sepsis in the ICU: A Retrospective Study.

Objective: The primary objectives of this study were to compare the characteristics of older and younger patients with sepsis and to analyze risk factors associated with 28-day and 90-day mortality in critically ill patients. Our study aimed to explore whether there are significant differences between sepsis patients in different age groups and whether these differences are related to the association between disease severity and mortality. Methods: We conducted a single-center, retrospective study of 5783 critically ill patients over 18 years of age from the Medical Information Mart for Intensive Care III database diagnosed with sepsis and admitted to the intensive care unit between 2008 and 2012. We performed a retrospective analysis, selected the Critical Care Medicine Information Mart III database, and collected data on patients with sepsis. We then collated and analyzed these data to compare differences in characteristics between older and younger patients and identify associated risk factors, which can help understand patient mortality. This approach leverages existing clinical data and avoids new experiments or data collection. Kaplan-Meier survival curve was used to assess 28-day and 90-day mortality, and a Cox proportional hazards regression model was used to evaluate the associated risk factors with 28-day and 90-day mortality. Results: Our study identified significant differences in mortality between older and younger patients with sepsis, finding that older patients had significantly higher mortality than younger patients. Furthermore, we successfully identified risk factors associated with mortality, results that have important implications for optimizing patient care and making clinical decisions. Of 5783 patients with sepsis, 2044 (35.3%) were younger than 60 years, and 3739 (64.7%) were aged 60 years or older. The 28-day mortality rate was 11.8% and 21.2% in the younger and older cohorts, respectively (P < .01). In the age-stratified analysis, the 28-day mortality was the highest in patients aged over 80 years (14.6% vs. 21.2% vs. 26.8%, P < .001). Factors associated with 28-day and 90-day mortality in patients with sepsis included age, weight, the need for mechanical ventilation, congestive heart failure, chronic pulmonary disease, malignancy, and Sequential Organ Failure Assessment score. Higher mortality in older patients with sepsis suggests the need for more aggressive treatment and monitoring. We also identified risk factors associated with mortality, helping to develop individualized treatment strategies. In addition, the different clinical characteristics of patients in different age groups emphasize the need for refined care pathways to meet their special needs. These results will help improve the treatment effect and quality of life of patients with sepsis. Conclusions: Our study fills the knowledge gap on the manifestations of sepsis patients in different age groups and helps medical staff better predict and manage disease progression in these two groups and provide personalized treatment. This lays the foundation for future in-depth research on age-related sepsis factors and is expected to improve patient survival and recovery rates. Older patients with sepsis had higher mortality rates and adverse outcomes. The mortality rate in patients with sepsis gradually increased with age. The importance of these findings is that they can help guide patient care and clinical decision-making, particularly when dealing with older and younger patients with sepsis, to improve treatment outcomes and reduce mortality. We would like to acknowledge that there are several limitations to the study, including the selectivity of the database and the retrospective nature, which preclude inference of causal relationships. In addition, some unconsidered variables may affect the results, and missing information in the data may also have an impact on the study. Future research could further explore these issues.This study highlights the critical role of age in sepsis patient outcomes and provides a strong basis for more sophisticated care and treatment. Our findings will help save more lives and improve patients' chances of recovery, which has profound implications for future research and clinical practice in the field of sepsis.

Intramodule pressure profiles and protein accumulation during tangential flow filtration.

Tangential flow filtration (TFF) through a 30 kDa nominal molecular weight cut-off (MWCO) ultrafiltration membrane is widely employed to concentrate purified monoclonal antibodies (mAbs) to levels required for their formulation into injectable biologics. While TFF has been used to remove casein from milk for cheese production for over 35 years, and in pharmaceutical manufacture of biotherapeutic proteins for 20 years, the rapid decline in filtration rate (i.e., flux) at high protein concentrations is a limitation that still needs to be addressed. This is particularly important for mAbs, many of which are 140-160 kDa immunoglobulin G (IgG) type proteins recovered at concentrations of 200 mg/mL or higher. This work reports the direct measurement of local transmembrane pressure drops and off-line confocal imaging of protein accumulation in stagnant regions on the surface of a 30 kDa regenerated cellulose membrane in a flat-sheet configuration widely used in manufacture of biotherapeutic proteins. These first-of-a-kind measurements using 150 kDa bovine IgG show that while axial pressure decreases by 58 psi across a process membrane cassette, the decrease in transmembrane pressure drop is constant at about 1.2 psi/cm along the 20.7 cm length of the membrane. Confocal laser scanning microscopy of the membrane surface at the completion of runs where retentate protein concentration exceeds 200 mg/mL, shows a 50 µm thick protein layer is uniformly deposited. The localized measurements made possible by the modified membrane system confirm the role of protein deposition on limiting ultrafiltration rate and indicate possible targets for improving membrane performance.

Comparative proteomic analysis of vancomycin-sensitive and vancomycin-intermediate resistant Staphylococcus aureus.

PURPOSE: The extensive use of vancomycin has led to the development of Staphylococcus aureus strains with varying degrees of resistance to vancomycin. The present study aimed to explore the molecular causes of vancomycin resistance by conducting a proteomics analysis of subcellular fractions isolated from vancomycin-intermediate resistant S. aureus (VISA) and vancomycin-sensitive S. aureus (VSSA) strains. METHODS: We conducted proteomics analysis of subcellular fractions isolated from 2 isogenic S. aureus strains: strain 11 (VSSA) and strain 11Y (VISA). We used an integrated quantitative proteomics approach assisted by bioinformatics analysis, and comprehensively investigated the proteome profile. Intensive bioinformatics analysis, including protein annotation, functional classification, functional enrichment, and functional enrichment-based cluster analysis, was used to annotate quantifiable targets. RESULTS: We identified 128 upregulated proteins and 21 downregulated proteins in strain 11Y as compared to strain 11. The largest group of differentially expressed proteins was composed of enzymatic proteins associated with metabolic and catalytic activity, which accounted for 32.1% and 50% of the total proteins, respectively. Some proteins were indispensable parts of the regulatory networks of S. aureus that were altered with vancomycin treatment, and these proteins were related to cell wall metabolism, cell adhesion, proteolysis, and pressure response. CONCLUSION: Our proteomics study revealed regulatory proteins associated with vancomycin resistance in S. aureus. Some of these proteins were involved in the regulation of cell metabolism and function, which provides potential targets for the development of strategies to manage vancomycin resistance in S. aureus.

Thrombospondin 1 and Reelin act through Vldlr to regulate cardiac growth and repair.

Adult mammalian cardiomyocytes have minimal cell cycle capacity, which leads to poor regeneration after cardiac injury such as myocardial infarction. Many positive regulators of cardiomyocyte cell cycle and cardioprotective signals have been identified, but extracellular signals that suppress cardiomyocyte proliferation are poorly understood. We profiled receptors enriched in postnatal cardiomyocytes, and found that very-low-density-lipoprotein receptor (Vldlr) inhibits neonatal cardiomyocyte cell cycle. Paradoxically, Reelin, the well-known Vldlr ligand, expressed in cardiac Schwann cells and lymphatic endothelial cells, promotes neonatal cardiomyocyte proliferation. Thrombospondin1 (TSP-1), another ligand of Vldlr highly expressed in adult heart, was then found to inhibit cardiomyocyte proliferation through Vldlr, and may contribute to Vldlr's overall repression on proliferation. Mechanistically, Rac1 and subsequent Yap phosphorylation and nucleus translocation mediate the regulation of the cardiomyocyte cell cycle by TSP-1/Reelin-Vldlr signaling. Importantly, Reln mutant neonatal mice displayed impaired cardiomyocyte proliferation and cardiac regeneration after apical resection, while cardiac-specific Thbs1 deletion and cardiomyocyte-specific Vldlr deletion promote cardiomyocyte proliferation and are cardioprotective after myocardial infarction. Our results identified a novel role of Vldlr in consolidating extracellular signals to regulate cardiomyocyte cell cycle activity and survival, and the overall suppressive TSP-1-Vldlr signal may contribute to the poor cardiac repair capacity of adult mammals.

2-Bromopalmitate inhibits malignant behaviors of HPSCC cells by hindering the membrane location of Ras protein.

Palmitoylation, which is mediated by protein acyltransferase (PAT) and performs important biological functions, is the only reversible lipid modification in organism. To study the effect of protein palmitoylation on hypopharyngeal squamous cell carcinoma (HPSCC), the expression levels of 23 PATs in tumor tissues of 8 HPSCC patients were determined, and high mRNA and protein levels of DHHC9 and DHHC15 were found. Subsequently, we investigated the effect of 2-bromopalmitate (2BP), a small-molecular inhibitor of protein palmitoylation, on the behavior of Fadu cells in vitro (50 µM) and in nude mouse xenograft models (50 µmol/kg), and found that 2BP suppressed the proliferation, invasion, and migration of Fadu cells without increasing cell apoptosis. Mechanistically, the effect of 2BP on the transduction of BMP, Wnt, Shh, and FGF signaling pathways was tested with qRT-PCR, and its drug target was explored with western blotting and acyl-biotinyl exchange assay. Our results showed that 2BP inhibited the transduction of the FGF/ERK signaling pathway. The palmitoylation level of Ras protein decreased after 2BP treatment, and its distribution in the cell membrane structure was reduced significantly. The findings of this work reveal that protein palmitoylation mediated by DHHC9 and DHHC15 may play important roles in the occurrence and development of HPSCC. 2BP is able to inhibit the malignant biological behaviors of HPSCC cells, possibly via hindering the palmitoylation and membrane location of Ras protein, which might, in turn, offer a low-toxicity anti-cancer drug for targeting the treatment of HPSCC.

Jianpi Qutan Fang induces anti-atherosclerosis and ameliorates endothelial cell injury in high-fat diet ratsan anti-inflammatory and inhibiting Janus kinase/signal transducer and activator of transcription signaling pathway.

OBJECTIVE: To investiage the possible mechanism underlying the effect of the Jianpi Qutan Fang (, JPQT) on Atherosclerosis (AS) which is the main pathological process of most cardiovascular diseases that affect millions of adults worldwide. METHODS: In the present study, rats were fed with a high-fat-diet (HFD) with vitamin D3 for 16 weeks and were orally administered atorvastatin treatment and different doses of JPQT. Histopathological changes and ultrastructural changes in the aorta were evaluated through hematoxylin-eosin staining and transmission electron microscopy (TEM), respectively. Suppressor of cytokine signaling 1 (SOCS1)/Janus kinase 1 (JAK1)/ signal transducer and activator of transcription 1 (STAT1) signaling pathways were detected through Western blotting. RESULTS: JPQT treatment decreased the lipid levels of triglyceride, low-density lipoprotein, and cholesterol, the inflammatory cytokine levels of interleukin 1 beta (IL-1ß), IL-6 and IL-8 in rat serum, but increased high-density lipoprotein and IL-10 serum levels. JPQT treatment ameliorated pathological changes in the aorta of AS model rats. Moreover, JPQT upregulated SOCS1 protein expression and down-regulated phosphorylated protein expression levels of p-JAK1 and p-STAT1. CONCLUSION: These results suggest that JPQT induces anti-atherosclerosis effects through anti-inflammatory and inhibiting JAK/STAT signaling pathways in HFD fed rats.

Extraction of Diatom DNA from Water Samples and Tissues.

Diatom testing is an essential auxiliary means in forensic practice to determine whether the corpse drowned in water and to infer the drowning location. Diatom testing is also an important research content in the field of the environment and plankton. The diatom molecular biology testing technology, which focuses on diatom DNA as the primary research object, is a new method of diatom testing. Diatom DNA extraction is the basis of diatom molecular testing. At present, the kits commonly used for diatom DNA extraction are expensive, which increases the cost of carrying out related research. Our laboratory improved the general whole blood genomic DNA rapid extraction kit and obtained a satisfactory diatom DNA extraction effect, thus providing an alternative economical and affordable DNA extraction solution based on glass beads for related research. The diatom DNA extracted using this protocol could satisfy many downstream applications, such as PCR and sequencing.