Inflammatory corpuscle AIM2 facilitates macrophage foam cell formation by inhibiting cholesterol efflux protein ABCA1.

The inflammatory corpuscle recombinant absents in melanoma 2 (AIM2) and cholesterol efflux protein ATP binding cassette transporter A1(ABCA1) have been reported to play opposing roles in atherosclerosis (AS) plaques. However, the relationship between AIM2 and ABCA1 remains unclear. In this study, we explored the potential connection between AIM2 and ABCA1 in the modulation of AS by bioinformatic analysis combined with in vitro experiments. The GEO database was used to obtain AS transcriptional profiling data; screen differentially expressed genes (DEGs) and construct a weighted gene co-expression network analysis (WGCNA) to obtain AS-related modules. Phorbol myristate acetate (PMA) was used to induce macrophage modelling in THP-1 cells, and ox-LDL was used to induce macrophage foam cell formation. The experiment was divided into Negative Control (NC) group, Model Control (MC) group, AIM2 overexpression + ox-LDL (OE AIM2 + ox-LDL) group, and AIM2 short hairpin RNA + ox-LDL (sh AIM2 + ox-LDL) group. The intracellular cholesterol efflux rate was detected by scintillation counting; high-performance liquid chromatography (HPLC) was used to detect intracellular cholesterol levels; apoptosis levels were detected by TUNEL kit; levels of inflammatory markers (IL-1ß, IL-18, ROS, and GSH) were detected by ELISA kits; and levels of AIM2 and ABCA1 proteins were detected by Western blot. Bioinformatic analysis revealed that the turquoise module correlated most strongly with AS, and AIM2 and ABCA1 were co-expressed in the turquoise module with a trend towards negative correlation. In vitro experiments demonstrated that AIM2 inhibited macrophage cholesterol efflux, resulting in increased intracellular cholesterol levels and foam cell formation. Moreover, AIM2 had a synergistic effect with ox-LDL, exacerbating macrophage oxidative stress and inflammatory response. Silencing AIM2 ameliorated the above conditions. Furthermore, the protein expression levels of AIM2 and ABCA1 were consistent with the bioinformatic analysis, showing a negative correlation. AIM2 inhibits ABCA1 expression, causing abnormal cholesterol metabolism in macrophages and ultimately leading to foam cell formation. Inhibiting AIM2 may reverse this process. Overall, our study suggests that AIM2 is a reliable anti-inflammatory therapeutic target for AS. Inhibiting AIM2 expression may reduce foam cell formation and, consequently, inhibit the progression of AS plaques.

2,5-Diketopiperazines From a Sponge-Derived Fungus Aspergillus sclerotiorum.

Three new 2,5-diketopiperazines, speramide C (1), 3,21-epi-taichunamide F (2), and 2-epi-amoenamide C (3), along with four known analogs (4-7), were obtained from the sponge-derived fungus Aspergillus sclerotiorum GDST-2013-0501 collected from the South China Sea. The chemical structures of new compounds were elucidated by analyzing NMR and MS spectroscopy data, and their absolute configurations were determined by electronic circular dichroism (ECD) calculations. Compound 1 represents the first prenylated indole alkaloid with an ethylene oxide ring at the isopentenyl side chain. Compound 4 displayed DNA topoisomerase I inhibitory activity and antibacterial activity against Staphylococcus epidermidis. The low cytotoxic or non-cytotoxic compound 4 displayed DNA topoisomerase I inhibitory activity, which could provide a starting point for the development of antitumor agents.

Transcriptome Analysis Reveals MFGE8-HAPLN3 Fusion as a Novel Biomarker in Triple-Negative Breast Cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) is a highly aggressive cancer with poor prognosis. The lack of effective targeted therapies for TNBC remains a profound clinical challenge. Fusion transcripts play critical roles in carcinogenesis and serve as valuable diagnostic and therapeutic targets in cancer. The present study aimed to identify novel fusion transcripts in TNBC. METHODS: We analyzed the RNA sequencing data of 360 TNBC samples to identify and filter fusion candidates through SOAPfuse and ChimeraScan analysis. The characteristics, including recurrence, fusion type, chromosomal localization, TNBC subgroup distribution, and clinicopathological correlations, were analyzed in all candidates. Furthermore, we selected the promising fusion transcript and predicted its fusion type and protein coding capacity. RESULTS: Using the RNA sequencing data, we identified 189 fusion transcripts in TNBC, among which 22 were recurrent fusions. Compared to para-tumor tissues, TNBC tumor tissues accumulated more fusion events, especially in high-grade tumors. Interestingly, these events were enriched at specific chromosomal loci, and the distribution pattern varied in different TNBC subtypes. The vast majority of fusion partners were discovered on chromosomes 1p, 11q, 19p, and 19q. Besides, fusion events mainly clustered on chromosome 11 in the immunomodulatory subtype and chromosome 19 in the luminal androgen receptor subtype of TNBC. Considering the tumor specificity and frameshift mutation, we selected MFGE8-HAPLN3 as a novel biomarker and further validated it in TNBC samples using PCR and Sanger sequencing. Further, we successfully identified three types of MFGE8-HAPLN3 (E6-E2, E5-E3, and E6-E3) and predicted the ORF of E6-E2, which could encode a protein of 712 amino acids, suggesting its critical role in TNBC. CONCLUSIONS: Improved bioinformatic stratification and comprehensive analysis identified the fusion transcript MFGE8-HAPLN3 as a novel biomarker with promising clinical application in the future.

Alternatone A, an Unusual Perylenequinone-Related Compound from a Soft-Coral-Derived Strain of the Fungus Alternaria alternata.

A novel perylenequinone-related compound, alternatone A (1), with an unprecedented tricyclo[6.3.1.02,7] dodecane skeleton, together with three known perylenequinones, altertoxin I (2), stemphyperylenol (3), and alterperylenol (4), was isolated from the soft-coral-derived fungus Alternaria alternata L3111'. Their structures including absolute configurations were elucidated on the basis of comprehensive spectroscopic analysis, electronic circular dichroism calculations, and X-ray diffraction data. Compound 4 showed cytotoxicity against A-549, HCT-116, and HeLa cell lines with IC50 values of 2.6, 2.4, and 3.1 µM, respectively. A possible biosynthetic pathway of 1 was proposed.

Effects of renal denervation on blood-pressure response to hemorrhagic shock in spontaneously hypertensive rats.

PURPOSE: Renal denervation (RD) has been demonstrated to be an effective approach to reduce blood pressure for those with resistant hypertension. Yet, we aimed to explore the effect and possible mechanism of RD on blood-pressure response to hemorrhagic shock in spontaneously hypertensive rats. METHODS: A total of 48 male spontaneously hypertensive rats were randomized to three groups: study group, sham-operation group and control group. RD was achieved by cutting off renal nerves and swabbing phenol on it. Ten weeks after RD, 8 rats in each group were sacrificed to collect the kidney and heart tissues. The remaining rats were subjected to an operation to induce hemorrhagic shock which would lead to 40% loss of total blood volume, and observed for 120 min. The serum concentration of norepinephrine was measured before and three weeks after RD. RESULTS: The blood-pressure and norepinephrine levels were reduced significantly after RD (p < 0.05). Systolic blood pressure and diastolic blood pressure of the surgery group were higher than those in the sham and control groups at 15, 30 and 45 min after hemorrhagic shock (p < 0.05), while no significant difference was observed at 60, 90 and 120 min (p > 0.05). Additionally, the beta-1 adrenergic receptor (ß1-AR) in the study group was significantly higher than those in the other two groups (p < 0.05) after hemorrhagic shock. CONCLUSION: This study demonstrated that RD could to some extent improve blood-pressure response to hemorrhagic shock in an established model of severe hemorrhagic shock in spontaneously hypertensive rats. The mechanism might be associated with up-regulation of ß1-AR.

The Effect of Taichi Practice on Attenuating Bone Mineral Density Loss: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

Objective: The purpose of this study was to determine the effects of practicing Taichi on attenuating bone mineral density (BMD) loss. Methods: Both electronic and manual searches were performed for randomized controlled trials (RCTs) examining Taichi for bone health. Two review authors independently performed study selection and data extraction according to inclusion criteria. A third party (Lin Luo) emerged to discuss with the two review authors and resolve a disagreement. Results: Twenty RCTs were found to meet the inclusion criteria and used for meta-analysis with a total effective sample of 1604. The aggregated results from this systematic review have shown significant benefits in favour of Taichi on BMD at lumbar spine (Standard Mean Difference, SMD) = 0.29; 95% CI 0.15 to 0.43; p < 0.0001), femur neck (SMD = 0.56; 95% CI 0.38 to 0.75; p < 0.00001), femur trochanter (SMD = 0.04; 95% CI 0.01 to 0.07; p = 0.007), total hip BMD (SMD = 0.46; 95% CI 0.16 to 0.76; p = 0.003). Conclusions: The aggregated results from this systematic review suggests that Taichi is effective on attenuating BMD loss at the regions of lumbar spine and proximal femur neck in special populations (e.g., older adults, perimenopausal and postmenopausal women, people with osteoarthritis, and cancer survivors). Researchers should further examine the effect of Taichi on the proximal femur trochanter and total hip so that a more definitive claim can be made regarding the beneficial effects for attenuating BMD loss in these musculoskeletal regions.

Application of reverse docking for target prediction of marine compounds with anti-tumor activity.

A large number of structures of anti-cancer drug targets have been solved and deposited to the protein data bank already. Identification of the targets for marine compounds with anti-tumor activity presents a challenge for marine natural products scientists. In this study, fast and efficient computational reverse docking was applied to predict the probable targeting proteins of the marine compounds with anti-tumor activity. Crystal structures of the proteins involved in tumor genesis, growth and metastasis were collected from PDB to construct the anti-tumor protein database (APD) for reverse docking. Two non-commercial docking programs, AutoDock Vina and LeDock, were used to perform the docking. Our results suggest that reverse docking is efficient for target fishing of compounds with known anti-tumor activities. In addition, the results show that performance of reverse docking using LeDock is superior to that using AutoDock Vina. Overall, reverse docking is a fast and efficient computational method to identify the probable target of the compounds with anti-tumor activities, and it can be complementary to the biological testing methods.

Pantoprazole blocks the JAK2/STAT3 pathway to alleviate skeletal muscle wasting in cancer cachexia by inhibiting inflammatory response.

OBJECTIVE: Cancer cachexia is often present in patients with advanced malignant tumors, and the subsequent body weight reduction results in poor quality of life. However, there has been no progress in developing effective clinical therapeutic strategies for skeletal muscle wasting in cancer cachexia. Herein, we explored the functions of pantoprazole on cancer cachexia skeletal muscle wasting. METHODS: The mouse colon adenocarcinoma cell line C26 was inoculated in the right forelimb of male BALB/C mice to establish a cancer cachexia model. The animals were treated with or without different concentrations of pantoprazole orally, and the body weight, tumor growth, spontaneous activity, and muscle functions were determined at various time points. Two weeks later, the levels of serum IL-6 and TNF-α, the mRNA levels of gastrocnemius JAK2 and STAT3, and the expression levels of p-JAK2, p-STAT3, Fbx32, and MuRF1 were examined with ELISA assay, qRT-PCR assay, and Western blotting, respectively. Further studies were performed to assess the levels of Fbx32 and MuRF1 expression and morphological changes. RESULTS: Pantoprazole can alleviate cancer cachexia-induced body weight reduction and inhibit skeletal muscle wasting in a dose-dependent manner. Our results indicated that pantoprazole treatment can decrease the levels of serum IL-6 and TNF-α (56.3% and 67.6%, respectively), and inhibit the activation of the JAK2/STAT3 signaling pathway. Moreover, the expression levels of MuRF1 and Fbx32 were also suppressed after pantoprazole treatment. CONCLUSION: Our findings suggested that pantoprazole can alleviate cancer cachexia skeletal muscle wasting by inhibiting the inflammatory response and blocking the JAK2/STAT3 or ubiquitin proteasome pathway.

Effects on heavy metal accumulation in freshwater fishes: species, tissues, and sizes.

Three fish species (Carassius auratus, Pelteobagrus fulvidraco, and Squaliobarbus curriculus) were collected from Xiang River near Changsha City, Southern China. The concentrations of heavy metals including arsenic (As), cadmium (Cd), copper (Cu), iron (Fe), manganese (Mn), lead (Pb), and zinc (Zn) in the muscle, gill, and liver of three species were determined by the inductively coupled plasma mass spectrometry method. One-way analysis of variance (ANOVA) was introduced to determine the significant variations (p < 0.05) of heavy metals. Livers were found to accumulate Cd and Cu due to the metallothionein proteins. High levels of Mn and Pb in the gills indicated that the main uptake pathway of these heavy metals was from the water. The carnivorous species, P. fulvidraco, was found to accumulate the highest levels of toxic elements (As, Cd, and Pb), while relatively high concentrations of nutrient elements (Cu, Fe, Mn, and Zn) were accumulated in omnivorous species (C. auratus and S. curriculus). According to the results of Pearson's correlation analysis, there were few significant relationships at p < 0.01 level between the concentrations of the analyzed elements and the fish sizes. The results of risk assessment indicated that exposure to the toxic heavy metals from fish muscle consumption posed no non-carcinogenic health risk to local inhabitants.

Simultaneous determination of haloanisoles and halophenols in water using in situ acylation combined with solid-phase microextraction with gas chromatography and mass spectrometry.

In this work, an in situ acylation combined with solid-phase microextraction coupled to gas chromatography and mass spectrometry method has been developed for simultaneously determining haloanisoles (2,4,6-trichloranisole, 2,4,6-tribromoanisole), and their direct precursors (2,4,6-trichlorophenol, 2,4,6-tribromophenol) and indirect precursors (2-chloropenol, 2,4-dichlorophenol, 2-bromophenol, 2,4-dibromophenol) in water. The key parameters for the solid-phase microextraction were determined by using Plackett-Burman screening and optimized by central composite optimization. Under optimal conditions, the eight compounds can be analyzed in a short time (33 min) with a strong linearity ranging from 2 to 200 ng/L (correlation coefficient greater than 0.996), showing good sensitivities with the limit of detection in a range of 0.23-0.91 ng/L and a limit of quantification of 0.77-3.03 ng/L, good repeatability (2.00-9.10%) and interday precision (1.67-11.3%). When environmental water samples were treated, the recoveries of target compounds were 75.5-127.3%, suggesting that the developed method could be applied in probing the origin of haloanisoles and monitoring halophenols and haloanisoles in natural waters at concentration levels of ng/L.

Aliskiren targets multiple systems to alleviate cancer cachexia.

To examine the effects of aliskiren, a small-molecule renin inhibitor, on cancer cachexia and to explore the underlying mechanisms. A cancer cachexia model was established by subcutaneously injecting C26 mouse colon carcinoma cells into isogenic BALB/c mice. Aliskiren was administered intragastrically [10 mg/kg body weight (BW)] on day 5 (as a preventive strategy, AP group) or on day 12 (as a therapeutic strategy, AT group) after C26 injection. Mice that received no C26 injection (healthy controls, HC group) or only C26 injection but not aliskiren (cancer, CA group) were used as controls. BW, tumor growth, whole body functions, and survival were monitored daily in half of the mice in each group, whereas serum, tumors, and gastrocnemius muscles were harvested from the other mice after sacrifice on day 20 for further analysis. Aliskiren significantly alleviated multiple cachexia­associated symptoms, including BW loss, tumor burden, muscle wasting, muscular dysfunction, and shortened survival. On the molecular level, aliskiren antagonized cachexia­induced activation of the renin­angiotensin system (RAS), systematic and muscular inflammation, oxidative stress, and autophagy­lysosome as well as ubiquitin­proteasome stimulation. In addition, early administration of aliskiren before cachexia development (AP group) resulted in more robust effects in alleviating cachexia or targeting underlying mechanisms than administration after cachexia development (AT group). Aliskiren exhibited potent anti­cachexia activities. These activities were achieved through the targeting of at least four mechanisms underlying cachexia development: RAS activation, increase in systematic inflammation, upregulation of oxidative stress, and stimulation of autophagy-lysosome pathway (ALP) and ubiquitin-proteasome pathway (UPP).

Economic burden of gastrointestinal cancer under the protection of the New Rural Cooperative Medical Scheme in a region of rural China with high incidence of oesophageal cancer: cross-sectional survey.

OBJECTIVE: To evaluate the financial burden of oesophageal cancer under the protection of the new Rural Cooperative Medical Scheme (NCMS) and to provide evidence and suggestions to policymakers in a high-incidence region in China. METHODS: We analysed inpatient claim data for oesophageal cancer, gastric cancer and colorectal cancer from 1 January to 31 December 2013. The data were extracted from the NCMS management system of Hua County, Henan Province, a typical high-risk region for oesophageal cancer in China. Cancer-specific health economic indicators were calculated to evaluate the financial burden under the protection of the local NCMS. RESULTS: The total cost of oesophageal cancer was 2.7-3.6 times higher than that of gastric cancer and colorectal cancer, respectively, due to high incidence of oesophageal cancer. For each hospitalisation to treat oesophageal cancer, the average total cost and out-of-pocket expenses after reimbursement equalled an entire year's gross domestic product per capita and per capita disposable income, respectively, for the local area. The average total cost per hospitalisation for oesophageal cancer increased monotonically with hospital level for surgical hospitalisations, and it increased more rapidly for non-surgical hospitalisations (from $301 to $2589, 860%) than for gastric cancer (from $289 to $1453, 503%) and colorectal cancer (from $359 to $1610, 448%). Vulnerable groups with less access to high-level hospitals were found in different gender and age groups. CONCLUSIONS: Oesophageal cancer imposes serious financial burdens on communities and patients' households in this high-incidence region, and no preferential policy from the local NCMS has been designed to address this issue. A special supportive policy should be developed on the basis of local disease profiles and population characteristics to alleviate the financial burden of populations at high risk for certain high-cost diseases.

Naphthalenones and Depsidones from a Sponge-Derived Strain of the Fungus Corynespora cassiicola.

Two new naphthalenones, corynenones A and B (1 and 2), and one new depsidone, corynesidone E (3), together with one known depsidone, corynesidone A (4) and two known diphenyl ethers, corynethers A (5) and B (6), were isolated from the sponge-derived fungus Corynespora cassiicola XS-20090I7. Their structures including absolute configurations were determined by spectroscopic data and electronic circular dichroism (ECD) spectra. Compounds 4 and 5 showed cytotoxicity against human promyelocytic leukemia HL-60 and human cervical carcinoma HeLa cell lines.

Combined treatment of carfilzomib and z-VAD-fmk inhibits skeletal proteolysis and apoptosis and ameliorates cancer cachexia.

The purpose of the study was to evaluate the therapeutic benefit of treatments with carfilzomib (CFZ) and z-VAD-fmk in a mouse model of cancer-induced cachexia. The model of cancer-associated cachexia was generated by injecting murine C26 adenocarcinoma cells into BALB/C mice. CFZ and z-VAD-fmk were administered individually or in combination at 5 and 12 days after inoculation. Changes in body weight, gastrocnemius muscle mass, tumor burden, spontaneous activity, survival, and metabolic profiles were noted. Also evaluated were the circulatory levels of renin and angiotensin II, and levels of apoptotic, proteolytic, and renin-angiotensin system-associated markers and transcription factor 2 (ATF2) in gastrocnemius muscle. The CFZ and z-VAD-fmk treatments were associated with less muscle wasting, reduced tumor burden, modulated metabolism, higher levels of glucose, albumin, and total proteins, and lower levels of triglyceride fatty acids, more spontaneous physical activity, and longer survival in C26-inoculated mice compared with PBS-treated cachectic mice. CFZ and z-VAD-fmk treatments resulted in higher levels of caspase-3 and BAX and lower level of BCL-XL in gastrocnemius muscles and altered the level of proteins in the renin-angiotensin system. The combined treatment administered 5 days after C26 inoculation was more effective than other regimens. Combined treatment with CFZ and z-VAD-fmk early in the development of cachexia was associated with signs of less proteolysis and apoptosis and less severe cachexia in a mouse model of cancer-induced cachexia.

Engineering a freestanding biomimetic cardiac patch using biodegradable poly(lactic-co-glycolic acid) (PLGA) and human embryonic stem cell-derived ventricular cardiomyocytes (hESC-VCMs).

Microgrooved thin PLGA film (≈30 µm) is successfully fabricated on a Teflon mold, which could be readily peeled off and is used for the construction of a biomimetic cardiac patch. The contraction of it is studied with optical mapping on transmembrane action potential. Our results suggest that steady-state contraction could be easily established on it under regular electrical stimuli. Besides, the biomimetic cardiac patch recapitulates the anisotropic electrophysiological feature of native cardiac tissue and is much more refractory to premature stimuli than the random one constructed with non-grooved PLGA film, as proved by the reduced incidence of arrhythmia. Considering the good biocompatibility of PLGA as demonstrated in our study and the biodegradability of it, our biomimetic cardiac patch may find applications in the treatment of myocardial infarction. Moreover, the Teflon mold could be applied in the fabrication of various scaffolds with fine features for other tissues.

[Prevalence and risk factors for hepatitis B in Hua County, Henan Province].

OBJECTIVE: To determine the epidemiological characteristics and associated risk factors of hepatitis B surface antigens (HBsAg) prevalence in rural areas of Hua County, Henan Province, and to provide scientific evidence for Hepatitis B prevention and control in local areas. METHODS: On the basis of an ongoing esophageal cancer cohort study in rural Anyang, a total of 5 104 subjects aged 25-65 years were clustered and selected from 5 targeted villages for this study in rural areas of Hua County, Henan Province. HBsAg was detected in their blood samples and a questionnaire was completed by all the subjects in a manner of one-on-one interview. All statistical analyses were conducted using SPSS for Windows version 13.0. RESULTS: Of the 5 104 studied subjects (overall participation rate: 92.05%), 5.17% were positive for HBsAg. The detection rate was significantly higher in males than in females (6.54% vs. 3.87%, P<0.001) and the highest detection rates were observed in the 25-29 and 55-59 years groups in both males and females. Multiple Logistic analyses showed unmarried status (OR=1.80, 95% CI: 1.00-3.25) and high frequency of sexual intercourse (Ptrend=0.049) were associated with higher the risk for hepatitis B virus (HBV) infection. CONCLUSION: The prevalence of HBsAg in rural Hua County, Henan Province, was slightly lower than the national average of the same time period. More attention should be attached to high risk groups of HBV infection in this population, i.e. males aged 25-29 years and 55-59 years and sexually active population. Immunization and health education projects against hepatitis B should be carried out in this population to further reduce the overall prevalence of hepatitis B.