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Objective: Salvia miltiorrhiza (SM) contains four major aqueous active ingredients, which have been isolated, purified and identified as danshensu (DSS), salvianolic acid A (Sal-A), salvianolic acid B (Sal-B) and protocatechuic aldehyde (PAL), A mixture of these four ingredients is called SABP. Although aqueous extract from Salvia Miltiorrhiza has been traditionally used to treat cardiovascular diseases, the efficacy and function of the optimal ratio of SABP in preventing and treating cardiovascular diseases remain unknown. This study aims to explore the anti-inflammatory mechanisms underlying the attenuation of atherosclerosis development by aqueous extract from Salvia Miltiorrhiza. Methods: Male ApoE-/- mice (6 weeks) were randomly allocated into three groups: the model group (Model), the SABP group (SABP), and the rosuvastatin calcium group (RC). Male C57BL/6 mice (6 weeks) were used as a control group. All mice were fed with an ordinary diet. After 8 weeks of treatment, the lipid profiles in serum and the lactate dehydrogenase (LDH) and creatine kinase (CK) in heart tissue were measured using an automatic biochemical analyzer. Alterations of the thoracic aorta and the heart were assessed using Hematoxylin and eosin staining. The protein expression of Toll-like receptor 4 (TLR4), TGF beta-activated kinase 1 (TAK1), nuclear factor kappa-B (NF-κB), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the heart tissue were determined though immunohistochemistry and western blotting analysis. Results: The serum low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), and total cholesterol (TC) levels were increased, and the high-density lipoprotein cholesterol (HDL-C) level was decreased in ApoE-/- mice. SABP significantly decreased serum lipid levels and improved histopathology in the thoracic aorta. In addition. SABP treatment inhibited the expression of TLR4, TAK1, NF-κB, IL-6 and TNF-α in the heart in ApoE-/- mice. The LDH and CK in the heart did not differ significantly among different groups, and the heart did not have obvious pathological changes. Conclusion: These findings indicated that SABP may exert an anti-atherosclerotic effect by lowering blood lipids and inhibiting inflammatory response via TLR4/ NF-κB signaling pathway.

Objective: Salvia Miltiorrhiza (SM) is a traditional Chinese medicine used clinically to treat cardiovascular diseases including atherosclerosis and myocardial infarction. Its therapeutic effect has been confirmed by many clinical and pharmacological studies. However, the optimal formulation of active ingredients in SM for treating cardiovascular diseases remains unclear. In this study, we determined the ratio of the optimal compatibility of SM ingredients DSS, Sal-A, Sal-B, and PAL (SABP）with a uniform and orthogonal optimized experimental design. In addition, we determined the anti-oxidation effect of SABP using Adventitial Fibroblasts (AFs). Methods: By using a combination of uniform and orthogonal designs, we determined the optimal formulation of aqueous extract from SM. MTT assay was used to determine the inhibitory effects of these 4 components of SM on the AFs, which were isolated and cultured from aorta. The reactive oxygen species (ROS) production in AFs was compared before and after SABP treatment. Results: The optimal formulation of these 4 aqueous extracts from SM were 150︰7︰300︰500, and their concentrations were S(1.5×10-4 mol/L), A(7×10-6 mol/L), B(3×10-4 mol/L), and P(5×10-4 mol/L). There were some synergies between these 4 components. Moreover, SABP decreased ROS production in AFs. Conclusion: These findings suggest that SABP inhibits the proliferation and oxidation stress in AFs. The present study provides a new evidence that the efficacy and function generated from optimal formulation of active ingredients in SM are better than lyophilized powder of SM.

Spermatogenesis is a highly regulated developmental process. During spermatogenesis, the replacement of histones by protamine plays an important role in repackaging the haploid genome of the sperm nuclear. The spermatozoan nucleus of the Decapoda is decondensed. Presently, many studies have proved that histones and histone modifications were retained in the nucleus or acrosome structure of the Decapoda spermatozoa. This review summarizes the dynamic characteristics of histones during spermatogenesis in Decapoda and helps clarify the mechanism of the decondensed chromatin structure.

Tissue phenotypic plasticity facilitates rapid adaptation of organisms to biotic and/or abiotic pressure. The reproductive capacity of honey bee workers (Apis mellifera) is plastic and responsive to pheromones produced by broods and the queen. Egg laying workers (ELWs), which could reactivate their ovaries and lay haploid eggs upon queen lost, have been commonly discussed from many aspects. However, it remains unclear whether midgut homeostasis in ELWs is affected during plastic changes. Here, we found that the expression of nutrition- and autophagy-related genes was up-regulated in the midguts of ELWs, compared with that in nurse workers (NWs) by RNA-sequencing. Furthermore, the area and number of autophagosomes were increased, along with significantly increased cell death in the midguts of ELWs. Moreover, cell cycle progression in the midguts of ELWs was increased compared with that in NWs. Consistent with the up-regulation of nutrition-related genes, the body and midgut sizes, and the number of intestinal proliferation cells of larvae reared with royal jelly (RJ) obviously increased more than those reared without RJ in vitro. Finally, cell proliferation was dramatically suppressed in the midguts of ELWs when autophagy was inhibited. Altogether, our data suggested that autophagy was induced and required to sustain cell proliferation in ELWs’ midguts, thereby revealing the critical role of autophagy played in the intestines during phenotypic plasticity changes.

Yellow slurry water is a kind of nutrient-rich wastewater of tofu. Firstly, the medium of yellow slurry was optimized. Then, APP40, APP60, and APP80 were obtained by sedimentation with different concentration of ethanol (40, 60, and 80%). The physicochemical properties and primary structures of the three polysaccharides were studied by high performance anion exchange chromatography (HPAEC), high performance gel filtration chromatography (HPGFC), scanning electron microscope (SEM), atomic force microscope (AFM), and Fourier transform infrared (FT-IR) spectrometer. Finally, the effects of three polysaccharides on antioxidation activity were studied. According to the experimental optimization the results, the biomass and the production of

Myocardial ischemia–reperfusion (MI/R) injury is characterized by iron deposition and reactive oxygen species production, which can induce ferroptosis. Ferroptosis has also been proposed to promote cardiomyocyte death. The current study sought to define the mechanism governing cardiomyocyte death in MI/R injury. An animal model of MI/R was established by ligation and perfusion of the left anterior descending coronary artery, and a cellular model of IR was constructed in cardiomyocytes. ChIP assay was then conducted to determine the interaction among USP22, SIRT1, p53, and SLC7A11. Loss- and gain-of-function assays were also conducted to determine the in vivo and in vitro roles of USP22, SIRT1, and SLC7A11. The infarct size and pathological changes of myocardial tissue were observed using TCC and hematoxylin–eosin staining, and the levels of cardiac function– and myocardial injury–related factors of rats were determined. Cardiomyocyte viability and apoptosis were evaluated in vitro, followed by detection of ferroptosis-related indicators (glutathione (GSH), reactive oxygen species, lipid peroxidation, and iron accumulation). USP22, SIRT1, and SLC7A11 expressions were found to be down-regulated, whereas p53 was highly expressed during MI/R injury. USP22, SIRT1, or SLC7A11 overexpression reduced the infarct size and ameliorated pathological conditions, cardiac function, as evidenced by reduced maximum pressure, ejection fraction, maximum pressure rate, and myocardial injury characterized by lower creatine phosphokinase and lactate dehydrogenase levels in vivo. Moreover, USP22, SIRT1, or SLC7A11 elevation contributed to enhanced cardiomyocyte viability and attenuated ferroptosis-induced cell death in vitro, accompanied by increased GSH levels, as well as decreased reactive oxygen species production, lipid peroxidation, and iron accumulation. Together, these results demonstrate that USP22 overexpression could inhibit ferroptosis-induced cardiomyocyte death to protect against MI/R injury via the SIRT1/p53/SLC7A11 association.

Salvia miltiorrhiza (SM) is a traditional Chinese medicine used clinically to treat cardiovascular diseases, including atherosclerosis and myocardial infarction. Its therapeutic effect has been confirmed by many clinical and pharmacological studies. However, the optimal formulation of active ingredients in SM for treating cardiovascular diseases remains unclear. In this study, we determined the ratio of the optimal compatibility of SM ingredients DSS, Sal-A, Sal-B, and PAL (SABP)with a uniform and orthogonal optimized experimental design. In addition, we determined the anti-oxidation effect of SABP using Adventitial Fibroblasts (AFs).By using a combination of uniform and orthogonal designs, we determined the optimal formulation of aqueous extract from SM. MTT assay was used to determine the inhibitory effects of these 4 components of SM on the AFs, which were isolated and cultured from the aorta. The reactive oxygen species (ROS) production in AFs was compared before and after SABP treatment.The optimal formulation of these 4 aqueous extracts from SM were 150 : 7 : 300 : 500, and their concentrations were S(1.5×10-4 mol/L), A(7×10-6 mol/L), B(3×10-4 mol/L), and P(5×10-4 mol/L). There were some synergies between these 4 components. Moreover, SABP decreased ROS production in AFs.These findings suggest that SABP inhibits the proliferation and oxidation stress in AFs. The present study provides new evidence that the efficacy and function generated from the optimal formulation of active ingredients in SM are better than lyophilized powder of SM.

Spermatogenesis in animals is the process by which male spermatogonia develop into mature spermatozoa. In most taxa, the process involves changes in the basic proteins associated with DNA. Somatic-type histones are partially or totally replaced by transition proteins, which in turn are replaced by protamines producing compact packaging of the genome. Sperm chromatin in the Chinese mitten crab (Eriocheir sinensis) has a noncompacted loosely arranged organization. However, its formation during spermatogenesis is not clear. In this study, a cDNA sequence encoding histone H2B was cloned by polymerase chain reaction amplification, and its recombinant protein was expressed and purified. Protein alignment studies demonstrated that this histone H2B had 80.80%, 95.12%, 80.16%, 91.87%, 81.75%, 77.78% and 99.19% identity with its counterparts in zebrafish, fruit fly, human, prawn, mouse, African clawed frog, and crayfish, respectively. Western blotting indicated that the recombinant protein could be recognized by an anti-H2B antibody and confirmed that histone H2B exists in sperm nuclei. Immunofluorescence demonstrated that histone H2B was present in the nuclei of spermatogonia, spermatocytes, spermatids, and mature spermatozoa. This is the first report that the mature sperm nucleus of E. sinensis contains histone H2B. This work complements a previous study of sperm histones of this species and provides a basis for further study of the noncondensed sperm nuclei of decapod crustaceans.

Eyestalk ablation is the most common procedure to induce gonadic maturation of Eriocheir sinensis. In addition to reproduction, other physiological and metabolic processes are also affected by the removal of X-organ sinus complex located in the eyestalk. In this study, we studied physiological and biochemical effects of eyestalk ablation of E. sinensis, and the results were as follows. The amylase activity of male crabs with unilateral or bilateral eyestalk ablaion was increased with the days increasing, and the maximal value was on the 20th day (P P

The global expansion of C4 plants during the Late Cenozoic is an important event in the Earth’s environmental-ecosystem evolution. Taking into account the nonglobal expansion of C4 plants, the CO2 concentration is not the only prerequisite for C4 expansion. The direct driving factors for C4 expansion may be associated with regional arid and/or warm seasonal precipitation and temperature variations. However, a large change in the CO2 concentration occurred, varying from 100∼300 to >2000 ppm during geological history; thus, it is necessary to understand the impact of such changes in the atmospheric CO2 concentration on the δ13C values of C3 and C4 plants during different geological periods. In this article, we analyzed the δ13C values for seven C4 and C3 plants grown under different CO2 concentrations for 10, 20, and 30 days. According to our study, the δ13C records for all the C3 and C4 plants show a rapid decrease followed by a slowly stable decreased trend with increasing CO2 concentration when the plants were grown for 10, 20, and 30 days. Comparing with the value averaging about −26 to −27 ‰ for C3 plants and about −12 ‰ for C4 plants, when the CO2 levels reach 20,000 ppm, all of the δ13C values of C4 plants become more negative, varying from −27.5 to −36.1 ‰, which fall into the range of δ13C values for C3 plants. Namely, the increase of carbon dioxide can limit the identification of C4 plants by carbon isotope composition. Thus, the expansion of C4 plants as reconstructed from the geological records of carbon isotopes possibly does not reflect the origin and expansion of C4 plants when atmospheric CO2 concentration was reduced to a threshold value, as previously thought. However, another possible explanation is that the atmospheric CO2 concentration was reduced to a threshold value from which the carbon isotope values of C3 and C4 plants can be distinguished. Therefore, we suggest that it is necessary to understand the δ13C range for C3 and C4 plants, to accurately evaluate the origin and expansion of C4 plants, for geological periods when a high CO2 concentration occurred.

Histones and histone phosphorylation play vital roles during animal spermatogenesis and spermatozoa maturation. The dynamic distribution of histones H2A and H4 and phosphorylated H2A and H4 at serine 1 (HS1ph) was explored in mammalian and Decapoda germ cells, with a special focus on the distribution of H2A, H4 and HS1ph between mouse condensed spermatozoa chromatin and crab non-condensed spermatozoa chromatin. The distribution of histone marks was also analysed in mature spermatozoa with different chromatin structures. Histone H2A and H4 marks were closely associated with the relatively loose chromatin structure in crab spermatozoa. The significant decrease in the HS1ph signal during spermatogenesis suggests that eliminating most of these epigenetic marks in the nucleusis closely associated with spermatozoa maturity.

During spermatogenesis in most animals, the basic proteins associated with DNA are continuously changing and somatic-typed histones are partly replaced by sperm-specific histones, which are then successively replaced by transition proteins and protamines. With the replacement of sperm nuclear basic proteins, nuclei progressively undergo chromatin condensation. The Chinese Mitten Crab (Eriocheir sinensis) is also known as the hairy crab or river crab (phylum Arthropoda, subphylum Crustacea, order Decapoda, and family Grapsidae). The spermatozoa of this species are aflagellate, and each has a spherical acrosome surrounded by a cup-shaped nucleus, peculiar to brachyurans. An interesting characteristic of the E. sinensis sperm nucleus is its lack of electron-dense chromatin. However, its formation is not clear. In this study, sequences encoding histones H3 and H4 were cloned by polymerase chain reaction amplification. Western blotting indicated that H3 and H4 existed in the sperm nuclei. Immunofluorescence and ultrastructural immunocytochemistry demonstrated that histones H3 and H4 were both present in the nuclei of spermatogonia, spermatocytes, spermatids and mature spermatozoa. The nuclear labeling density of histone H4 decreased in sperm nuclei, while histone H3 labeling was not changed significantly. Quantitative real-time PCR showed that the mRNA expression levels of histones H3 and H4 were higher at mitotic and meiotic stages than in later spermiogenesis. Our study demonstrates that the mature sperm nuclei of E. sinensis contain histones H3 and H4. This is the first report that the mature sperm nucleus of E. sinensis contains histones H3 and H4. This finding extends the study of sperm histones of E. sinensis and provides some basic data for exploring how decapod crustaceans form uncondensed sperm chromatin.