Integrating Network Pharmacology and In Vitro Experimental Verification Revealing Bushenhuoxue Recipe Against Intrauterine Adhesions via PI3K-AKT Signaling Pathway.

We aimed to investigate therapeutic effect of Bushenhuoxue recipe in intrauterine adhesions (IUA) and explore the underlying molecular mechanism via integrating network pharmacology and in vitro experimental verification. The active compounds and gene targets of Bushenhuoxue recipe were screened in the TCMSP database and the IUA-related genes were identified using GeneCards database by the keyword "Intrauterine adhesions". Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were conducted to reveal the underlying molecular mechanism of Bushenhuoxue recipe treating IUA. T-HESC cells were inducted to fibrotic state using TGF-ß1 of 10 ng/ml concentration treating for 24 h. RT-qPCR or western blot was used to demonstrate the expression levels of fibrosis markers (COL1A1 and α-SMA) and KEGG pathway markers. Cell counting kit-8 (CCK8) assay was performed to illustrate the cell viability of endometrial stromal cell. The treatment of Bushenhuoxue recipe could significantly inhibit the proliferation and fibrosis of endometrial stromal cells. We obtained a total of 169 no-repeat ingredients of Bushenhuoxue recipe and 3044 corresponding targets. After taking intersection with 4230 no-repeat IUA-related genes, a total of 83 target genes related to both Bushenhuoxue recipe and IUA were finally identified. KEGG analysis found that PI3K-AKT signaling pathway might be the key pathway. Further experiment revealed that PI3K-AKT signaling pathway was significantly activated in endometrial stromal cells of fibrotic state and the treatment of Bushenhuoxue recipe could inhibit the PI3K-AKT signaling pathway. Further rescue assay demonstrated that Bushenhuoxue recipe suppressed the proliferation and fibrosis of endometrial stromal cells via PI3K-AKT signaling pathway. Bushenhuoxue recipe suppresses the proliferation and fibrosis of endometrial stromal cells via PI3K-AKT signaling pathway, eventually inhibiting the progression of IUA.

AGEs promote atherosclerosis by increasing LDL transcytosis across endothelial cells via RAGE/NF-κB/Caveolin-1 pathway.

OBJECTIVE: To elucidate the mechanism whereby advanced glycation end products (AGEs) accelerate atherosclerosis (AS) and to explore novel therapeutic strategies for atherosclerotic cardiovascular disease. METHODS AND RESULTS: The effect of AGEs on low-density lipoprotein (LDL) transcytosis across endothelial cells (ECs) was assessed using an in vitro model of LDL transcytosis. We observed that AGEs activated the receptor for advanced glycation end products (RAGE) on the surface of ECs and consequently upregulated Caveolin-1, which in turn increased caveolae-mediated LDL transcytosis and accelerated AS progression. Our molecular assessment revealed that AGEs activate the RAGE-NF-κB signaling, which then recruits the NF-κB subunit p65 to the RAGE promoter and consequently enhances RAGE transcription, thereby forming a positive feedback loop between the NF-κB signaling and RAGE expression. Increased NF-κB signaling ultimately upregulated Caveolin-1, promoting LDL transcytosis, and inhibition of RAGE suppressed AGE-induced LDL transcytosis. In ApoE-/- mice on a high-fat diet, atherosclerotic plaque formation was accelerated by AGEs but suppressed by EC-specific knockdown of RAGE. CONCLUSION: AGEs accelerate the development of diabetes-related AS by increasing the LDL transcytosis in ECs through the activation of the RAGE/NF-κB/Caveolin-1 axis, which may be targeted to prevent or treat diabetic macrovascular complications.

Tracking Carbon Flows in China's Iron and Steel Industry.

Accurately tracking carbon flows is the first step toward reducing the climate impacts of the iron and steel industry (ISI), which is still lacking in China. In this study, we track carbon flows from coal/mineral mines to end steel users by coupling the cross-process material and energy flow model, point-based emission inventory, and interprovincial trade matrices. In 2020, ISI emitted 2288 Tg of CO2 equivalent (CO2eq, including CH4 and CO2), 96% of which came from energy use and 4% from raw material decomposition. Often overlooked off-gas use and CH4 leakage in coal mines account for 25% of life-cycle emissions. Due to limited scrap resources and a high proportion of pig iron feed, the life-cycle emission intensity of the electric arc furnace (EAF) (1.15 t CO2eq/t steel) is slightly lower than the basic oxygen furnace (BOF) (1.58 t CO2eq/t steel) in China. In addition, over 49% of producer-based emissions are driven by interprovincial coal/coke/steel trade. In particular, nearly all user-based emissions in Zhejiang and Beijing are transferred to steelmaking bases. Therefore, we highlight the need for life-cycle and spatial shifts in user-side carbon management.

Diagnostic Value of the Combination of Ultrasonographic Optic Nerve Sheath Diameter and Width of Crural Cistern with Respect to the Intracranial Pressure in Patients Treated with Decompressive Craniotomy.

BACKGROUND: The monitoring of intracranial pressure (ICP) and detection of increased ICP are crucial because such increases may cause secondary brain injury and a poor prognosis. Although numerous ultrasound parameters, including optic nerve sheath diameter (ONSD), width of the crural cistern (WCC), and the flow velocities of the central retinal artery and middle cerebral artery, can be measured in patients after hemicraniectomy, researchers have yet to determine which of these is better for evaluating ICP. This study aimed to analyze the correlation between ICP and ultrasound parameters and investigate the best noninvasive estimator of ICP. METHODS: This observational study enrolled 50 patients with brain injury after hemicraniectomy from January 2021 to December 2021. All patients underwent invasive ICP monitoring with microsensor, transcranial, and ocular ultrasound postoperatively. We measured the ONSD including the dura mater (ONSDI), the ONSD excluding the dura mater, the optic nerve diameter (OND), the eyeball transverse diameter (ETD), the WCC, and the flow velocities in the central retinal artery and middle cerebral artery. Then, we calculated the ONSDI-OND (the difference between ONSDI and OND) and ONSDI/ETD (the ratio of ONSDI to ETD). Patients were divided into a normal ICP group (n = 35) and an increased ICP group (≥ 20 mm Hg, n = 15) according to the ICP measurements. Correlations were then assessed between the values of the ultrasound parameters and ICP. RESULTS: The ONSDI, ONSDI-OND, and ONSDI/ETD were positively associated with ICP (r = 0.455, 0.482, 0.423 and p = 0.001, < 0.001, 0.002, respectively), whereas the WCC was negatively associated with ICP (r = - 0.586, p < 0.001). The WCC showed the highest predictive power for increased ICP (area under the receiver operating characteristic curve [AUC] = 0.904), whereas the ONSDI-OND and ONSDI also presented with acceptable predictive power among the ONSD-related parameters (AUC = 0.831, 0.803, respectively). The cutoff values for increased ICP prediction for ONSDI, ONSDI-OND, and WCC were 6.29, 3.03, and 3.68 mm, respectively. The AUC of the combination of ONSDI-OND and WCC was 0.952 (95% confidence interval 0.896-1.0, p < 0.001). CONCLUSIONS: The ONSDI, ONSDI-OND, and WCC were correlated with ICP and had acceptable accuracy levels in estimating ICP in patients after hemicraniectomy. Furthermore, WCC showed a higher diagnostic value than ONSD-related parameters, and the combination of ONSDI-OND and WCC was a satisfactory predictor of increased ICP.

An NF-κB-responsive long noncoding RNA, PINT, regulates TNF-α gene transcription by scaffolding p65 and EZH2.

Long noncoding RNAs (lncRNAs) are central regulators of the inflammatory response and play an important role in inflammatory diseases. PINT has been reported to be involved in embryonic development and tumorigenesis. However, the potential functions of PINT in the innate immune system are largely unknown. Here, we revealed the transcriptional regulation of inflammatory genes by PINT, whose expression is primarily dependent on the NF-κB signaling pathway in human and mouse macrophage and intestinal epithelial cell lines. Functionally, PINT selectively regulates the expression of TNF-α in basal and LPS-stimulated cells. Mechanistically, PINT acts as a modular scaffold of p65 and EZH2 to coordinate their localization and specify their binding to the target genes. Further, a high expression level of PINT was detected in intestinal mucosal tissues from patients with ulcerative colitis (UC). Together, these findings demonstrate that PINT acts as an activator of inflammatory responses, highlighting the importance of this lncRNA as a potential therapeutic target in infectious diseases and inflammatory diseases.

Crystal Structure and Noncovalent Interactions of Heterocyclic Energetic Molecules.

Nitrogen-rich heterocyclic compounds are important heterocyclic substances with extensive future applications for energetic materials due to their outstanding density and excellent physicochemical properties. However, the weak intermolecular interactions of these compounds are not clear, which severely limits their widespread application. Three nitrogen-rich heterocyclic compounds were chosen to detect their molecular geometry, stacking mode and intermolecular interactions by crystal structure, Hirshfeld surface, RDG and ESP. The results show that all atoms in each molecule are coplanar and that the stacking mode of the three crystals is a planar layer style. A large amount of inter- and intramolecular interaction exists in the three crystals. All principal types of intermolecular contacts in the three crystals are N···H interactions and they account for 40.9%, 38.9% and 32.9%, respectively. Hydrogen bonding, vdW interactions and steric effects in Crystal c are stronger than in Crystals a and b. The negative ESPs all concentrate on the nitrogen atoms in the three molecules. This work is expected to benefit the crystal engineering of heterocyclic energetic materials.

Pollution characteristics of soil heavy metals around two typical copper mining and beneficiation enterprises in Northwest China.

In order to investigate the situation of heavy metal pollution in the heavy metal industry in Gansu Province, a large copper mining province, two large and typical copper mining and beneficiation enterprises with differences in topographic features, climatic conditions, and soil types were selected as the target of this study based on similar ore types and beneficiation processes. Around these two enterprises, geochemical baselines of the six heavy metals were established, while the degree of local soil heavy metal pollution and potential hazards to humans were assessed based on statistical analysis, single-factor and multi-factor index analysis, and health risk evaluation models. In addition, Spearman's correlation analysis and hierarchical cluster analysis were used to explore the intrinsic association between each heavy metal in the two mining industries to reveal the pattern of soil heavy metal pollution in the copper mining and beneficiation industry and to propose targeted measures to improve and prevent soil heavy metal pollution. The results showed that the heavy metal pollution in the soil around Shengxi Mining Co., Ltd. of Subei County (SX enterprise) was higher than that around Yangba Copper Co., Ltd. of Gansu Province (YB enterprise), but the two enterprises had similar patterns of pollution, with an overall medium level of pollution. The carcinogenic and non-carcinogenic risks for children and adults were within acceptable limits for both enterprises. Besides, the correlation between the different heavy metals to similarity in their sources of contamination and the different degrees of association between the soil heavy metals of the two enterprises due to their environmental characteristics.

A Long Noncoding RNA, Antisense IL-7, Promotes Inflammatory Gene Transcription through Facilitating Histone Acetylation and Switch/Sucrose Nonfermentable Chromatin Remodeling.

Long noncoding RNAs are important regulators of gene expression in innate immune responses. Antisense IL-7 (IL-7-AS) is a newly discovered long noncoding RNA in human and mouse that has been reported to regulate the expression of IL-6. However, the potential function of IL-7-AS in innate immune system is not fully understood. In this study, we found that the expression of IL-7-AS is primarily dependent on the NF-κB and MAPK signaling pathways in macrophages and intestinal epithelial cells. Functionally, IL-7-AS promotes the expression of several inflammatory genes, including CCL2, CCL5, CCL7, and IL-6, in cells in response to LPS. Specifically, IL-7-AS physically interacts with p300 to regulate histone acetylation levels around the promoter regions of these gene loci. Moreover, IL-7-AS and p300 complex modulate the assembly of SWI/SNF complex to the promoters. IL-7-AS regulates chemotaxis activity of monocytes to intestine epithelial cells with involvement of CCL2. Therefore, our data indicate a new promoting role for NF-κB/MAPK-responsive IL-7-AS in the transcriptional regulation of inflammatory genes in the innate immune system although modulation of histone acetylation around the promoters of related genes.

Age-related changes of human serum Sirtuin6 in adults.

BACKGROUND: Aging is a natural life process and with an aging population, age-related diseases (e.g. type 2 diabetes mellitus (T2DM), atherosclerosis-based cardiovascular diseases) are the primary mortality cause in older adults. Telomerase is often used as an aging biomarker. Detection and characterization of novel biomarkers can help in a more specific and sensitive identification of a person's aging status. Also, this could help in age-related diseases early prevent, ultimately prolonging the population's life span. Sirtuin 6 (Sirt6) - a member of the Sirtuins NAD+-dependent histone deacetylases family - is mainly intracellularly expressed, and is reported to be involved in the regulation of aging and aging-related diseases. Whether serum Sirt6 is correlated with aging and could be used as an aging biomarker is unknown. In the present study, we aimed to investigate the age-related Sirt6 changes in the serum of human adults. METHODS: Participants were divided into three groups according to age: 20-30 years (Young); 45-55 years (Middle-aged); and ≥ 70 years (Old). The Sirt6 and telomerase serum concentrations were determined by ELISA. The Sirt6 and human telomerase reverse transcriptase (hTERT) expression in vessels from amputated human lower limbs were analyzed using real-time quantitative PCR (RT-qPCR) and immunohistochemical staining. The relationships between variables were evaluated by Pearson correlation analysis. RESULTS: The Sirt6 and telomerase serum levels reduced with an increase in age. A similar tendency was observed for Sirt6 and hTERT in the vessel. Serum levels of Sirt6 were higher in females compared with males. Pearson's regression analysis revealed that the Sirt6 serum level positively correlated with telomerase (r = 0.5743) and both were significantly negatively correlated with age (r = - 0.5830 and r = - 0.5993, respectively). CONCLUSIONS: We reported a negative correlation between serum Sirt6 concentration and aging in human beings. Therefore, the Sirt6 serum level is a potential sex-specific aging marker.

The NF-κB-Responsive Long Noncoding RNA FIRRE Regulates Posttranscriptional Regulation of Inflammatory Gene Expression through Interacting with hnRNPU.

Long noncoding RNAs, a newly identified class of noncoding RNAs, are important regulators of gene expression in innate immunity. We report in this study that the transcription of FIRRE, a conserved long noncoding RNA between humans and mice, is controlled by NF-κB signaling in macrophages and intestinal epithelial cells. Functionally, FIRRE appears to positively regulate the expression of several inflammatory genes in macrophages or intestinal epithelial cells in response to LPS stimulation via posttranscriptional mechanisms. Specifically, FIRRE physically interacts with heterogeneous nuclear ribonucleoproteins U, regulating the stability of mRNAs of selected inflammatory genes through targeting the AU-rich elements of their mRNAs in cells following LPS stimulation. Therefore, our data indicate a new regulatory role for NF-κB-responsive FIRRE in the posttranscriptional regulation of inflammatory genes in the innate immune system.

Autocrine motility factor receptor promotes the malignancy of glioblastoma by regulating cell migration and invasion.

OBJECTIVE: One of the important causes of death in cancer patients is malignant metastasis, invasion, and metastasis of tumor cells. Metastasis is also the most basic physiological characteristics and pathogenesis of various tumors. Previously published studies have suggested that autocrine motor factor receptor (AMFR) is the key regulator of tumor cell migration and invasion. Meanwhile, AMFR is highly expressed in esophageal tumors, gastrointestinal tumors, and bladder cancer, and it is also involved in its pathogenesis. However, the role of AMFR in glioblastoma has not been reported. METHODS: In order to study the role of AMFR in the cell migration and invasion of glioblastoma, AMFR was silenced using siRNA and overexpressed using cDNA. Immunoblotting analysis and real-time quantitative polymerase chain reaction (PCR) were employed to assess the expression of AMFR. We conducted wound healing assay, cell migration assay, and tumorsphere formation assay to detect the invasion and metastatic ability of glioblastoma. RESULTS: This study found that the level of AMFR expression was significantly correlated with the malignant degree of glioma tissue in clinic samples. AMFR silencing decreased cell migration and invasion of LN229. Overexpression of AMFR significantly increased cell migration and invasion of U251. CONCLUSION: This study suggests that AMFR could be used as a therapeutic strategy for the clinical treatment of glioblastoma.

Comparison of an interpretable extreme gradient boosting model and an artificial neural network model for prediction of severe acute pancreatitis.

INTRODUCTION: Acute pancreatitis (AP) that progresses to persistent organ failure is referred to as severe acute pancreatitis (SAP). It is a condition associated with a relatively high mortality. A prediction model that would facilitate early recognition of patients at risk for SAP is crucial for improvement of patient prognosis. OBJECTIVES: The aim of this study was to evaluate the accuracy of extreme gradient boosting (XGBoost) and artificial neural network (ANN) models for predicting SAP. PATIENTS AND METHODS: A total of 648 patients with AP were enrolled. XGBoost and ANN models were developed and validated in the training (519 patients) and test sets (129 patients). The accuracy and predictive performance of the XGBoost and ANN models were evaluated using both the area under the receiver operating characteristic curves (AUCs) and the area under the precision­recall curves (AUC­PRs). RESULTS: A total of 15 variables were selected for model construction through a univariable analysis. The AUCs of the XGBoost and ANN models in 5­fold cross­validation of the training set were 0.92 (95% CI, 0.87-0.97) and 0.86 (95% CI, 0.78-0.92), respectively, whereas the AUCs for the test set were 0.93 (95% CI, 0.85-1) and 0.87 (95% CI, 0.79-0.96), respectively. The XGBoost model outperformed the ANN model in terms of both diagnostic accuracy and AUC­PR. Individual predictions of the XGBoost model were explained using a local interpretable model­agnostic explanation plot. CONCLUSIONS: An interpretable XGBoost model showed better discriminatory efficiency for predicting SAP than the ANN model, and could be used in clinical practice to identify patients at risk for SAP.

Siglec-5 as a novel receptor mediates endothelial cells oxLDL transcytosis to promote atherosclerosis.

BACKGROUND: Excessive subendothelial retention of oxidized low-density lipoprotein (oxLDL) and subsequent oxLDL engulfment by macrophages leads to the formation of foam cells and the development of atherosclerosis. Our previous study showed that the plasma level of sialic acid-binding immunoglobulin-like lectin 5 (Siglec-5) was a novel biomarker for the prognosis of atherosclerosis in diabetic patients. However, the role and underlying mechanisms of Siglec-5 in atherosclerosis have not been elucidated. METHODS: The interaction between oxLDL and Siglec-5 was detected by fluorescence colocalization and coimmunoprecipitation. The effect of oxLDL on Siglec-5 expression was detected in endothelial cells and macrophages, and the effect of Siglec-5 on oxLDL transcytosis and uptake was investigated. Siglec-5 was overexpressed in mice using recombinant adeno-associated virus vector serotype 9 (rAAV9-Siglec-5) to evaluate the effect of Siglec-5 on oxLDL uptake and atherogenesis in vivo. In addition, the effects of Siglec-5 antibodies and soluble Siglec-5 proteins on oxLDL transcytosis and uptake and their role in atherogenesis were investigated in vivo and in vitro. RESULTS: We found that oxLDL interacted with Siglec-5 and that oxLDL stimulated the expression of Siglec-5. Siglec-5 promotes the transcytosis and uptake of oxLDL, while both anti-Siglec-5 antibodies and soluble Siglec-5 protein attenuated oxLDL transcytosis and uptake. Interestingly, overexpression of Siglec-5 by recombinant adeno-associated viral vector serotype 9 (rAAV9-Siglec-5) promoted the retention of oxLDL in the aorta of C57BL/6 mice. Moreover, overexpression of Siglec-5 significantly accelerated the formation of atherosclerotic lesions in Apoe-/- mice. Moreover, both anti-Siglec-5 antibodies and soluble Siglec-5 protein significantly alleviated the retention of oxLDL in the aorta of rAAV9-Siglec-5-transfected C57BL/6 mice and the formation of atherosclerotic plaques in rAAV9-Siglec-5-transfected Apoe-/- mice. CONCLUSION: Our results suggested that Siglec-5 was a novel receptor that mediated oxLDL transcytosis and promoted the formation of foam cells. Interventions that inhibit the interaction between oxLDL and Siglec-5, including anti-Siglec-5 antibody or soluble Siglec-5 protein treatment, may provide novel therapeutic strategies in treating atherosclerosis.

Nanoscale chestnut soil pore characteristics changes induced by non-growing season precipitation in a temperate grassland.

Changes in precipitation patterns during the non-growing season can affect soil moisture storage in temperate grasslands. However, there is a lack of comprehensive understanding regarding how these changes influence microscale soil pore characteristics and nutrient cycling in the context of climate change. Therefore, we carried out a 3-year artificial precipitation experiment during the non-growing season, along with N2 adsorption experiments of soil pore distribution and surveys of soil nutrient content. The aim was to clarify the influence of non-growing season precipitation variations on nanoscale soil pore characteristics and explore the potential correlations of the soil physicochemical properties. The results showed that: (1) The precipitation sheltering treatment during the non-growing season led to a significant 9.80 % increase in soil porosity at the 0-15 cm depth compared to the control. (2) Compared to the control, alterations in non-growing season precipitation (both increase and sheltering treatments) led to a significant increase in soil specific surface area (SSA), with an average increase of 23.2 %. Additionally, soil micropores, mesopores, macropores, and total pore volume (PV) increased by an average of 24.2 %, 14.0 %, 30.1 %, and 23.1 %, respectively. (3) Significant correlations were observed between soil microscale pore characteristics and soil C, soil organic matter (SOM), C: N ratio, and available P (AP). Redundancy analysis showed that soil microscale pore characteristics effectively accounted for the variations in soil nutrients with an explanatory degree of 94.23 %. (4) Influence pathways analysis by structural equation modeling indicated that dramatic variability in non-growing season precipitation promoted increases in mesopore and macropore volume, as well as the transformation of mesopores into macropores, thereby facilitating soil carbon accumulation. Our study suggests that soil microscale pore characteristics, acquired through adsorption experiments, assist in elucidating these potential synergistic mechanisms among physicochemical properties under varying non-growing season precipitation patterns. Given the escalating impacts of climate change, our findings provide novel insights and evidence for the assessment of climate change impacts in temperate arid grassland ecosystems.

The suitability of isotopic methods in identifying water sources of a shallow-rooted herbaceous plant in a desert steppe.

Isotopic methodologies have gained prominence in investigating the composition of plant water sources; however, concerns regarding their suitability and reliability in diverse environments have emerged in recent years. This study presents a comparative analysis of root, soil, and liquid water (precipitation, dew, and groundwater) samples obtained from a desert steppe using isotope ratio infrared spectrometry (IRIS) and isotope ratio mass spectrometry (IRMS). The objective was to evaluate the applicability of these techniques in discerning the water sources of Stipa breviflora, a shallow-rooted herbaceous plant species. Additionally, we explored the root water uptake characteristics and water use strategy of S. breviflora. Our findings indicate that the IRIS method had more enriched values of D compared to the IRMS method across all samples, while no discernible pattern was observed for 18O. Notably, the differences observed among all samples exceeded the instruments' accuracies. Moreover, an unexpected occurrence was noted, whereby both D and 18O values in the root water were more enriched than in any of the considered water sources, rendering identification of the plant water sources unattainable. By conducting a re-analysis of more refined soil layer samples, we discovered that S. breviflora exhibits the ability to absorb and utilize water sources in close proximity to the soil surface. It further suggested that the shallow-rooted herbaceous plants in desert steppes can exploit small rainfalls, frequently overlooked in their ecological importance. Considering the distinctive soil and plant characteristics of desert steppes, we recommend adopting IRMS methods in conjunction with refined surface soil sampling for isotopic analysis aiming to identify water sources of shallow-rooted herbaceous plants. This study provides novel insights into assessing the suitability of isotopic techniques for analyzing plant water sources, while enhancing our understanding of water use strategies and environmental adaptation mechanisms employed by shallow-rooted herbaceous plants within xerophytic grassland ecosystems.

Relationship between Cholesterol-Related Lipids and Severe Acute Pancreatitis: From Bench to Bedside.

It is well known that hypercholesterolemia in the body has pro-inflammatory effects through the formation of inflammasomes and augmentation of TLR (Toll-like receptor) signaling, which gives rise to cardiovascular disease and neurodegenerative diseases. However, the interaction between cholesterol-related lipids and acute pancreatitis (AP) has not yet been summarized before. This hinders the consensus on the existence and clinical importance of cholesterol-associated AP. This review focuses on the possible interaction between AP and cholesterol-related lipids, which include total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and apolipoprotein (Apo) A1, from the bench to the bedside. With a higher serum level of total cholesterol, LDL-C is associated with the severity of AP, while the persistent inflammation of AP is allied with a decrease in serum levels of cholesterol-related lipids. Therefore, an interaction between cholesterol-related lipids and AP is postulated. Cholesterol-related lipids should be recommended as risk factors and early predictors for measuring the severity of AP. Cholesterol-lowering drugs may play a role in the treatment and prevention of AP with hypercholesterolemia.

Scavenger receptor a mediates glycated LDL transcytosis across endothelial cells to promote atherosclerosis.

Glycated low-density lipoprotein (G-LDL) is an established proatherosclerotic factor, but the mechanism is not completely understood. In vitro, we evaluated the uptake and transcytosis rates of N-LDL and G-LDL in endothelial cells and the uptake and transcytosis rates of G-LDL were much higher than those of N-LDL. Then, using small interfering RNAs, the receptor mediating G-LDL uptake and transcytosis was screened among eight candidate receptors, and the mechanism of the receptor regulation was thoroughly examined. We discovered that scavenger receptor A (SR-A) knockdown dramatically decreased the uptake and transcytosis rates of G-LDL. Additionally, endothelial cells with overexpressed SR-A had enhanced G-LDL uptake and transcytosis. In vivo, G-LDL was injected in the tail vein of ApoE-/- mice to investigate whether G-LDL affects atherosclerotic plaque formation. Compared with the injection of N-LDL, the injection of G-LDL accelerated atherosclerotic plaque formation in ApoE-/- mice, which was ameliorated by endothelial cells specific SR-A knockdown. Together, our results provide the first demonstration that the transcytosis of G-LDL across endothelial cells is much faster than that of N-LDL and SR-A is the major type of receptor responsible for G-LDL binding and transcytosis across endothelial cells.

PrPC interacts with potassium channel tetramerization domain containing 1 (KCTD1) protein through the PrP(51-136) region containing octapeptide repeats.

To identify molecular interaction partners of the cellular prion protein (PrP(C)), we applied a yeast two-hybrid screen on a bovine brain cDNA expression library and identified the potassium channel tetramerization domain containing 1 (KCTD1) as a PrP(C) interacting protein. Deletion mapping showed that PrP(C) specifically binds KCTD1 through the unstructured PrP(51-136) region. We further confirmed the interaction between PrP(C) and KCDT1 protein by co-immunoprecipitation in vivo and by a biosensor assay in vitro. Interestingly, the binding of an insertion mutant PrP(8OR) to KCTD1 is higher than that of wild-type PrP(C), suggesting an important role for an unstructured region harboring octapeptide repeats in the KCTD1-PrP(C) interaction. Our results identify a novel PrP(C)-interacting protein and suggest a new approach to investigating the unidentified physiological cellular function of PrP(C).

Expression, purification, and secondary structure characterization of recombinant KCTD1.

Potassium channel tetramerization domain containing 1 (KCTD1) contains a BTB domain, which can facilitate protein-protein interactions that may be involved in the regulation of signaling pathways. Here we describe an expression and purification system that can provide a significant amount of recombinant KCTD1 from Escherichia coli. The cDNA encoding human KCTD1 was amplified and cloned into the expression vector pET-30a(+). The recombinant protein was expressed in E. coli BL21(DE3) cells and subsequently purified using affinity chromatography. To confirm that KCTD1 was correctly expressed and folded, the molecular weight and conformation were analyzed using mass spectroscopy, Western blot, and circular dichroism. Optimizing KCTD1 expression and investigating its secondary structure will provide valuable information for future structural and functional studies of KCTD1 and KCTD family proteins.

Poly[di-µ(3)-chlorido-di-µ(2)-chlorido-{µ(4)-N,N,N',N'-tetra-kis-[(diphenyl-phosphan-yl)meth-yl]benzene-1,4-diamine-κ(4)P:P':P'':P'''}tetra-copper(II)].

In the title complex, [Cu(4)Cl(4)(C(58)H(52)N(2)P(4))](n), four Cu(II) atoms are held together via two doubly bridging and two triply bridging chlorides, forming a stair-like Cu(4)Cl(4) core having crystallographically imposed inversion symmetry, while the benzene-1,4-diamine ligand (with a crystallographic inversion center at the centroid) acts in a tetra-dentate coordination mode, bridging two adjacent Cu(4)Cl(4) cores, resulting in a chain along the a-axis direction. One Cu atom has a distorted tetra-hedral geometry, coordinated by one P atom, one µ(2)-Cl and two µ(3)-Cl atoms, while the second Cu atom adopts a trigonal geometry, coordinated by one P atom, one µ(2)-Cl and one µ(3)-Cl atoms.