The plant extract PNS mitigates atherosclerosis via promoting Nrf2-mediated inhibition of ferroptosis through reducing USP2-mediated Keap1 deubiquitination.

BACKGROUND AND PURPOSE: Atherosclerosis is the basis of cardiovascular disease. Ferroptosis is a form of programmed cell death characterized by lipid peroxidation, which contributes to atherogenesis. The plant extract PNS (Panax notoginseng saponins), containing the main active ingredients of Panax notoginseng, exhibits anti-atherogenic properties. Herein, we determined whether PNS and its major components could attenuate atherosclerosis by suppressing ferroptosis and revealed the underlying mechanism(s). EXPERIMENTAL APPROACH: The anti-atherogenic effects of PNS and their association with inhibition of ferroptosis was determined in apoE-/- mice. In vitro, the anti-ferroptotic effect and mechanism(s) of PNS components were demonstrated in the presence of ferroptosis inducers. Expression of ferroptosis markers and the ubiquitination of Keap1 were evaluated in USP2-/- macrophages. Finally, the anti-atherogenic effect of USP2 knockout was determined by using USP2-/- mice treated with high-fat diet (HFD) and AAV-PCSK9. KEY RESULTS: PNS inhibited ferroptosis and atherosclerosis in vivo. PNS suppressed ferroptosis and ferroptosis-aggravated foam cell formation and inflammation in vitro. Mechanistically, PNS and its components activated Nrf2 by antagonizing Keap1, which was attributed to the inhibition of USP2 expression. USP2 knockout antagonized ferroptosis and ferroptosis-aggravated foam cell formation and inflammation, thus mitigating atherosclerosis. USP2 knockout abolished inhibitory effects of PNS on foam cell formation and inflammation in vitro. CONCLUSION AND IMPLICATIONS: PNS reduced USP2-mediated Keap1 de-ubiquitination and promoted Keap1 degradation, thereby activating Nrf2, improving iron metabolism and reducing lipid peroxidation, thus contributing to an anti-atherosclerotic outcome. Our study revealed the mechanism(s) underlying inhibition of ferroptosis and atherosclerosis by PNS.

Grassland degradation affected vegetation carbon density but not soil carbon density.

BACKGROUND: With the profound changes in the global climate, the issue of grassland degradation is becoming increasingly prominent. Grassland degradation poses a severe threat to the carbon cycle and carbon storage within grassland ecosystems. Additionally, it will adversely affect the sustainability of food production. The grassland ecosystem in the northwest region of Liaoning Province, China, is particularly vulnerable due to factors such as erosion from the northern Horqin Sandy Land, persistent arid climate, and issues related to overgrazing and mismanagement of grassland. The degradation issue is especially pronounced in this ecological environment. However, previous research on the carbon density of degraded grasslands in Northeast China has predominantly focused on Inner Mongolia, neglecting the impact on the grasslands in the northwest of Liaoning Province. Therefore, this experiment aims to assess the influence of grassland degradation intensity on the vegetation and soil carbon density in the northwest of Liaoning Province. The objective is to investigate the changes in grassland vegetation and soil carbon density resulting from different degrees of grassland degradation. METHODOLOGY: This study focuses on the carbon density of grasslands at different degrees of degradation in the northwest of Liaoning Province, exploring the variations in vegetation and soil carbon density under different levels of degradation. This experiment employed field sampling techniques to establish 100 × 100 m plots in grasslands exhibiting varying degrees of degradation. Six replications of 100 × 100 m plots per degradation intensity were sampled. Vegetation and soil samples were collected for analysis of carbon density. RESULTS: The results indicate that in the context of grassland degradation, there is a significant reduction in vegetation carbon density. Furthermore, it was found that root carbon density is the primary contributor to vegetation carbon density. In comparison to mildly degraded grasslands, moderately and severely degraded grasslands experience a reduction in vegetation carbon density by 25.6% and 52.6%, respectively. However, with regard to the impact of grassland degradation on soil carbon density, it was observed that while grassland degradation leads to a slight decrease in soil carbon density, there is no significant change in soil carbon density in the short term under the influence of grassland degradation. CONCLUSIONS: Therefore, grassland degradation has exerted a negative impact on aboveground vegetation carbon density, reducing the carbon storage of above-ground vegetation in grasslands. However, there was no significant effect on grassland soil carbon density.

MaiJiTong granule attenuates atherosclerosis by reducing ferroptosis via activating STAT6-mediated inhibition of DMT1 and SOCS1/p53 pathways in LDLR-/- mice.

BACKGROUND AND PURPOSE: Atherosclerosis is the primary pathological basis of cardiovascular disease. Ferroptosis is a regulated form of cell death, a process of lipid peroxidation driven by iron, which can initiate and promote atherosclerosis. STAT6 is a signal transducer that shows a potential role in regulating ferroptosis, but, the exact role in ferroptosis during atherogenesis remains unclear. The Traditional Chinese Medicine Maijitong granule (MJT) is used for treating cardiovascular disease and shows a potential inhibitory effect on ferroptosis. However, the antiatherogenic effect and the underlying mechanism remain unclear. In this study, we determined the role of STAT6 in ferroptosis during atherogenesis, investigated the antiatherogenic effect of MJT, and determined whether its antiatherogenic effect was dependent on the inhibition of ferroptosis. METHODS: 8-week-old male LDLR-/- mice were fed a high-fat diet (HFD) at 1st and 10th week, respectively, to assess the preventive and therapeutic effects of MJT on atherosclerosis and ferroptosis. Simultaneously, the anti-ferroptotic effects and mechanism of MJT were determined by evaluating the expression of genes responsible for lipid peroxidation and iron metabolism. Subsequently, we reanalyzed microarray data in the GSE28117 obtained from cells after STAT6 knockdown or overexpression and analyzed the correlation between STAT6 and ferroptosis. Finally, the STAT6-/- mice were fed HFD and injected with AAV-PCSK9 to validate the role of STAT6 in ferroptosis during atherogenesis and revealed the antiatherogenic and anti-ferroptotic effect of MJT. RESULTS: MJT attenuated atherosclerosis by reducing plaque lesion area and enhancing plaque stability in both preventive and therapeutic groups. MJT reduced inflammation via suppressing inflammatory cytokines and inhibited foam cell formation by lowering the LDL level and promoting ABCA1/G1-mediated lipid efflux. MJT ameliorated the ferroptosis by reducing lipid peroxidation and iron dysregulation during atherogenesis. Mechanistically, STAT6 negatively regulated ferroptosis by transcriptionally suppressing SOCS1/p53 and DMT1 pathways. MJT suppressed the DMT1 and SOCS1/p53 via stimulating STAT6 phosphorylation. In addition, STAT6 knockout exacerbated atherosclerosis and ferroptosis, which abolished the antiatherogenic and anti-ferroptotic effects of MJT. CONCLUSION: STAT6 acts as a negative regulator of ferroptosis and atherosclerosis via transcriptionally suppressing DMT1 and SOCS1 expression and MJT attenuates atherosclerosis and ferroptosis by activating the STAT6-mediated inhibition of DMT1 and SOCS1/p53 pathways, which indicated that STAT6 acts a novel promising therapeutic target to ameliorate atherosclerosis by inhibiting ferroptosis and MJT can serve as a new therapy for atherosclerosis treatment.

GLSP and GLSP-derived triterpenes attenuate atherosclerosis and aortic calcification by stimulating ABCA1/G1-mediated macrophage cholesterol efflux and inactivating RUNX2-mediated VSMC osteogenesis.

Background and Purpose: Atherosclerosis is the main pathophysiological foundation of cardiovascular disease, which was caused by inflammation and lipid metabolism disorder, along with vascular calcification. Aortic calcification leads to reduced plaque stability and eventually causes plaque rupture which leads to cardiovascular events. Presently, the drug to treat aortic calcification remains not to be available. Ganoderma lucidum spore powder (GLSP) is from Ganoderma lucidum which is a Traditional Chinese Medicine with the homology of medicine and food. It has multiple pharmacological effects, but no research on aortic calcification during atherosclerosis was performed. This study investigated the effects of GLSP on atherosclerosis and aortic calcification and revealed the underlying mechanism. Methods: In vivo, 8-week-aged male LDLR-/- mice were fed a high-fat diet to induce atherosclerosis along with aortic calcification. Simultaneously, the mice were treated with GLSP at the first week of HFD feeding to determine the protection against early and advanced atherosclerosis. Subsequently, the mice tissues were collected to evaluate the effects of GLSP on atherosclerosis, and aortic calcification, and to reveal the underlying mechanism. In vitro, we determined the major components of GLSP triterpenes by HPLC, and subsequently assessed the protective effects of these main active components on lipid metabolism, inflammation, and calcification in RAW264.7 and HASMC cells. Results: We observed GLSP attenuated plaque area and aortic calcification in the mice with early and advanced atherosclerosis. GLSP reduced the number of foam cells by improving ABCA1/G1-mediated cholesterol efflux in macrophages. In addition, GLSP protected against the aortic endothelium activation. Moreover, GLSP inhibited aortic calcification by inactivating RUNX2-mediated osteogenesis in HASMCs. Furthermore, we determined the major components of GLSP triterpenes, including Ganoderic acid A, Ganoderic acid B, Ganoderic acid C6, Ganoderic acid G, and Ganodermanontriol, and found that these triterpenes promoted ABCA1/G1-mediated cholesterol efflux and inhibited inflammation in macrophage, and inactivated RUNX2-mediated osteogenesis in VSMC. Conclusions: This study demonstrates that GLSP attenuates atherosclerosis and aortic calcification by improving ABCA1/G1-mediated cholesterol efflux and inactivating RUNX2-mediated osteogenesis in LDLR-/- mice. GLSP may be a potential drug candidate for the treatment of atherosclerosis and vascular calcification.

Exploring Ganweikang Tablet as a Candidate Drug for NAFLD Through Network Pharmacology Analysis and Experimental Validation.

Nonalcoholic fatty liver disease (NAFLD) is defined as liver disease in which more than 5% of hepatocytes are steatotic with little or no alcohol consumption. NAFLD includes benign nonalcoholic fatty liver (NAFL) and nonalcoholic steatohepatitis (NASH). Importantly, NASH is an advanced progression of NAFL and is characterized by steatosis, hepatocyte ballooning, lobular inflammation, and fibrosis. However, to date, no drugs specifically targeting NAFLD have been approved by the FDA. Therefore, a new drug or strategy for NAFLD treatment is necessary. However, the pathogenesis of NAFLD is complex and no single-target drugs have achieved the desired results. Noticeably, traditional Chinese medicine formulations are a complex system with multiple components, multiple targets, and synergistic effects between components. The Ganweikang tablet is a compound formula based on traditional Chinese medicine theory and clinical experience. In this study, network pharmacology analysis indicates Ganweikang tablet as a candidate for NAFLD treatment. Furthermore, we evaluated the therapeutic effects of Ganweikang tablet on the NAFL and NASH and tried to clarify the underlying molecular mechanisms in animal models and cell experiments. As expected, Ganweikang tablet was found to improve NAFL and NASH by modulating inflammation, apoptosis, and fatty acid oxidation by inhibiting NFκB, caspase-8, and activating PPARα, which not only indicates that Ganweikang tablet as a drug candidate but also provides a theoretical basis of Ganweikang tablet for the treatment of NAFL and NASH.

The Induction of Endothelial Autophagy and Its Role in the Development of Atherosclerosis.

Increasing attention is now being paid to the important role played by autophagic flux in maintaining normal blood vessel walls. Endothelial cell dysfunction initiates the development of atherosclerosis. In the endothelium, a variety of critical triggers ranging from shear stress to circulating blood lipids promote autophagy. Furthermore, emerging evidence links autophagy to a range of important physiological functions such as redox homeostasis, lipid metabolism, and the secretion of vasomodulatory substances that determine the life and death of endothelial cells. Thus, the promotion of autophagy in endothelial cells may have the potential for treating atherosclerosis. This paper reviews the role of endothelial cells in the pathogenesis of atherosclerosis and explores the molecular mechanisms involved in atherosclerosis development.

Multifunctional mandibles of ants: Variation in gripping behavior facilitated by specific microstructures and kinematics.

The elongated mandibles of certain ant species are dexterous grippers that can output a wide range of forces as needed for various tasks. Our combined experimental and theoretical research reveals the multifunctionality of the mandibles of Harpegnathos venator that is facilitated by specific microstructures and characteristic kinematics. First, we found that H. venator can pull off a spider's (Heteropoda venatoria) leg by closing its long mandibles. We observed that the ant usually clamps the spider's leg using the distal or middle part of its mandibles. In contrast, the ant can grip its egg with the proximal parts of its mandibles without causing damage. Our results showed that the spider's legs are always fractured at the coxa-trochanteral joint. Second, we found that the force required to fracture the spider's leg can be up to 500 times the ant's body weight. On the other hand, the maximum force can be controlled to less than 2×10- 6N while gripping an egg. By combining microstructure imaging, kinematic tracking and mathematical modeling, we uncovered that the sharp teeth and dense bristles on the internal side of the mandibles determine the high adhesion force, while the concave teeth and biaxial rotation of the mandibles facilitate gentle gripping. We validated our findings by constructing an artificial mandible pair. This work expands the knowledge of the physiological multifunctionality in ant mandibles, and provides novel ways to reveal the multifunctionality in insect appendages by applying the tools of mechanical analysis and related experimental devices.

Genome sequencing and annotation of Kocuria turfanensis HO-9042, to promote explorating high-salt resistance associated gene resources.

Kocuria turfanensis HO-9042 which was cultured from the extreme environment in China is a gram-positive, aerobic, non-motile and coccoid bacterium. In order to dissect genetic determinants of strong resistance to high salt, whole genome of K. turfanensis was sequenced. Then effective genetic engineering could be performed to promote gene resources application, including plant breeding in high-salt soil.

Complete genome sequence of Rufibacter tibetensis strain 1351, a radiation-resistant bacterium from Tibet plateau.

Rufibacter tibetensis strain 1351, isolated from the soil of the Tibet plateau of China, belongs to the family of Cytophagaceae. It is a red-pigmented, gram-negative, strictly aerobic and rod-shaped bacterium and shows resistance to UV radiation. Here, we report its complete genome sequence, which can help us find the key genes of the carotenoid biosynthesis and resistance to UV radiation.

[Prevalence of Angiostrongylus cantonensis infection in snails for sale in Fuzhou and Xiamen].

OBJECTIVE: To investigate the prevalence of Angiostrongylus cantonensis infection in snails for sale from Fuzhou and Xiamen of Fujian Province. METHODS: During 2009-2012, two markets and five restaurants from each city were selected as surveillance sites. A. cantonensis infection rate in Pomacea canaliculata, Bellamya aeruginosa and Cipangopaludina cathayensis was examined two times per month. More than 50 P. canaliculata or C. cathayensis, and about 500 g B. aeruginosa were collected in each site. A. cantonensis larvae infection was determined by lung-microscopy in P. canaliculata, and by tissue homogenate method in C. cathayensis and B. aeruginosa, respectively. RESULTS: In markets, a total of 5,744 P. canaliculata were collected, and the infection rate of A. cantonensis larvae was 13.8% (753/5 744) with the lowest prevalence in 2009 (8.4%, 28/334) and the highest one in 2011 (16.7%, 361/2,160). The overall infection rate of A. cantonensis showed an increasing trend over the past years (P < 0.05). In restaurants, 879 P. canaliculata snails were examined in Xiamen City, and the infection rate was 12.8% (877/6 879). No significant difference was found among years (P > 0.05). A. cantonensis larvae were found from P. canaliculata for sale in different seasons with no statistical difference (P > 0.05). 19,843 B. aeruginosa snails were collected in markets from the two cities, and the infection rate was 0.2% (31/19 843). The infection rate was highest in 2011 (0.3%, 16/5,953) and lowest in 2010 (0.04%, 2/4,706). All the 361 C. cathayensis snails were negative. CONCLUSION: A. cantonensis larvae are found in P. canaliculata and B. aeruginosa from markets and restaurants of Fuzhou and Xiamen in different seasons during the years.

[Prevalent trend of the infection of soil-transmitted nematodes in Fujian Province].

OBJECTIVE: To understand the endemic situation of soil-transmitted nematodes in Fujian Province. METHODS: According to the national guidelines, the survey spots were determined by clustered random sampling in nine prefectures (cities) of Fujian Province from 2007 to 2009. Residents of 3 years old and above were investigated. The eggs of roundworm, hookworm and whipworm in feces were checked by Kato-Katz method. Eggs per gram (EPG) in feces were calculated. The gender, age and education status of the investigated subjects were recorded. The results were evaluated in comparison to those in 1992 and 2003. RESULTS: Altogether 93 833 residents in 610 villages of 184 towns from 49 counties were investigated. The overall infection rate of soil-transmitted nematodes was 10.14% (9 511/93 833), decreased by 86.88% and 71.84% compared to that in 1992 and 2003, respectively. The prevalence of roundworm, hookworm and whipworm was 1.32%(1 234/93 833), 7.31% (6 863/93 833) and 1.73% (1 622/93 833), respectively. The average EPG of roundworm, hookworm and whipworm was 9 556, 526 and 156, respectively. The prevalence in males and females was 9.48% (4 385/46 246) and 10.77% (5 126/47 587), respectively, with a statistical difference (chi2 = 42.84, P < 0.01). There was also a statistical difference among the age groups (chi2 = 1 626, P < 0.01). The higher education level of the people, the lower prevalence (chi2 = 1 107, P < 0.01). CONCLUSION: The prevalence of soil-transmitted nematodes is on a downward trend in Fujian Province, but remained high in the underdeveloped areas. The hookworm infection rate is higher than the average of the nation.

[Surveillance on the prevalence of soil-transmitted nematode infection in Fujian in 2006-2010].

OBJECTIVE: To analyze the prevalent trend of soil-transmitted nematode infection in Fujian Province during the past 5 year surveillance and evaluate the control effect. METHODS: From 2006 to 2010, fecal samples of the inhabitants of 3 years old and above were collected every November and examined for intestinal helminth eggs by the modified Kato s thick smear technique at the 2 surveillance sites: Punan village of Zhangzhou and Gushan village of Shaowu. Cellophane tapes were used to detect pinworm eggs for children aged 3-12. Soil samples were also collected from vegetable field, lavatory, courtyard and kitchen of 20 randomly selected families (in 2 villages) each with stool egg-positive findings and examined for ascaris eggs by a modified saturated sodium nitrate floatation method. RESULTS: The prevalence of soil-transmitted nematode infection at the surveillance sites decreased from 45.3% (946/2087) in 2006 to 15.1% (226/1494) in 2010, with a reduction of 66.6%. Among the infected subjects, hookworm infection occupied 75%-85%, while ascaris or trichuris infections each accounted for less than 10%. In terms of infection intensity, 65.2%-85.5% of the hookworm infection was light, and majority of the infected subjects were farmers. The pinworm prevalence in children were still high although it had dropped down from 46.1% (140/304) in 2006 to 29.8% (36/121) in 2010, declined by 35.4%. In the 5 years, totally 400 soil samples from 100 families were examined and 21 samples were found ascaris egg positive with viable eggs in only one sample. CONCLUSION: The 5 year surveillance reveals a decreasing trend of the soil-transmitted nematode prevalence but shows a relatively high hookworm infection rate in the population and pinworm infection in children.

[Survey on the current status of important human parasitic infections in Fujian province].

OBJECTIVE: To understand the prevalence of important human parasitic diseases in Fujian province. METHODS: According to the national guidelines, clustered random sampling was made for the survey. Modified Kato-Katz thick smear was applied to examine helminth eggs for soil-transmitted nematodes and Clonorchis sinensis. Cellophane and swab technique was applied for detecting Enterobius vermicularis eggs in children under 12. Questionnairing, skin test, serological test and eosinophil examination were carried out for Paragonimus infection. Questionnairing and serological test were used for Taenia infection and cysticercosis. Retrospective investigation was conducted with the data on outpatient and hospitalized cases with parasitic diseases. RESULTS: 45,736 people at 73 spots of 24 counties were investigated and 14 species of parasites were found. The overall infection rate of intestinal helminth infections was 36.15%. The prevalence of Ascaris lumbricoides, hookworms and Trichuris trichiura was 9.55%, 19.73% and 11.48% respectively. The infection rate of Enterobius vermicularis and Clonorchis sinensis was 22.06% and 0.60% respectively. The positive rate of skin test and serological test among skin test positives for Paragonimus infection was 7.07% and 9.23% respectively. The serology positive rate for cysticercosis was 2.28%, only 3 cases of taenia infection were found from 16371 persons investigated. Human infections with Angiostrongylus cantonensis, Diphyllobothrium latus, Taenia saginata, and Gnathostoma spinigerum were found for the first time in the Province. Metorchis orientalis and Echinostoma aegyptiaca infections in human were first recorded. CONCLUSION: The overall prevalence of soil-transmitted nematodes has decreased significantly in the province, although still highly prevalent in social-economically less developed areas. Meanwhile, attention should be paid to the food-borne parasitic infections in human population.