TFPI from erythroblasts drives heme production in central macrophages promoting erythropoiesis in polycythemia.

Bleeding and thrombosis are known as common complications of polycythemia for a long time. However, the role of coagulation system in erythropoiesis is unclear. Here, we discover that an anticoagulant protein tissue factor pathway inhibitor (TFPI) plays an essential role in erythropoiesis via the control of heme biosynthesis in central macrophages. TFPI levels are elevated in erythroblasts of human erythroblastic islands with JAK2V617F mutation and hypoxia condition. Erythroid lineage-specific knockout TFPI results in impaired erythropoiesis through decreasing ferrochelatase expression and heme biosynthesis in central macrophages. Mechanistically, the TFPI interacts with thrombomodulin to promote the downstream ERK1/2-GATA1 signaling pathway to induce heme biosynthesis in central macrophages. Furthermore, TFPI blockade impairs human erythropoiesis in vitro, and normalizes the erythroid compartment in mice with polycythemia. These results show that erythroblast-derived TFPI plays an important role in the regulation of erythropoiesis and reveal an interplay between erythroblasts and central macrophages.

Evaluation of aristolochic acid Ι nephrotoxicity in mice via 1H NMR quantitative metabolomics and network pharmacology approaches.

Background: Although many studies have shown that herbs containing aristolochic acids can treat various human diseases, AAΙ in particular has been implicated as a nephrotoxic agent. Methods and results: Here, we detail the nephrotoxic effect of AAΙ via an approach that integrated 1H NMR-based metabonomics and network pharmacology. Our findings revealed renal injury in mice after the administration of AAΙ. Metabolomic data confirmed significant differences among the renal metabolic profiles of control and model groups, with significant reductions in 12 differential metabolites relevant to 23 metabolic pathways. Among them, there were seven important metabolic pathways: arginine and proline metabolism; glycine, serine, and threonine metabolism; taurine and hypotaurine metabolism; ascorbate and aldehyde glycolate metabolism; pentose and glucosinolate interconversion; alanine, aspartate, and glutamate metabolism; and glyoxylate and dicarboxylic acid metabolism. Relevant genes, namely, nitric oxide synthase 1 (NOS1), pyrroline-5-carboxylate reductase 1 (PYCR1), nitric oxide synthase 3 (NOS3) and glutamic oxaloacetic transaminase 2 (GOT2), were highlighted via network pharmacology and molecular docking techniques. Quantitative real-time PCR findings revealed that AAI administration significantly downregulated GOT2 and NOS3 and significantly upregulated NOS1 and PYCR1 expression and thus influenced the metabolism of arginine and proline. Conclusion: This work provides a meaningful insight for the mechanism of AAΙ renal injury.

TET3 as a non-invasive screening tool for the detection of fibrosis in patients with chronic liver disease.

Ten-eleven translocation protein 3 (TET3) is one of the key enzymes in DNA demethylation which can be expressed in liver tissues. However, the clinical value of TET3 for diagnosis and treatment of chronic liver disease have not been reported previously. We investigated the diagnostic accuracy of serum TET3 as a non-invasive screening tool for liver fibrosis. 212 patients with chronic liver disease from were enrolled in this study. Enzyme-linked immunosorbent assay was used to measure the serum levels of TET3. Receiver operating characteristics (ROC) were determined to examine the diagnostic accuracy of TET3 and combination model for diagnosis fibrosis. Serum TET3 level in fibrosis cases was significantly higher than that in non-fibrosis and controls, respectively. The areas under the ROC curve of the TET3 and fibrosis-4 index for liver fibrosis were 0.863 and 0.813, and 0.916 and 0.957 for liver cirrhosis. The combination of TET3 and fibrosis-4 index had a highly promising positive predictive value for detecting liver fibrosis and cirrhosis different stages of (93.5% and 100%) as compared with each diagnostic tool alone. TET3 is related to the development of liver fibrosis and cirrhosis. The TET3-fibrosis-4 model enhances discriminatory power and represents a promising non-invasive tool for the diagnosis and screening of liver fibrosis.

Metabonomics and 16S rRNA gene sequencing to study the therapeutic mechanism of Danggui Sini decoction on collagen-induced rheumatoid arthritis rats with Cold Bi syndrome.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by persistent joint inflammation. The development of rheumatoid arthritis is directly correlated with the disturbance of gut microbiome and its metabolites. RA can be effectively treated with the Danggui Sini decoction (DSD), a Traditional Chinese medicine (TCM) prescription from the Treatise on Febrile Diseases. Further research is needed to clarify the precise mechanism of DSD in the treatment of RA. In this study, 1H NMR metabonomics and 16 S rRNA gene sequencing techniques were used to clarify the intervention of DSD on CIA-induced RA. The results of 1H NMR metabolomics of feces revealed that five metabolites (alanine, glucose, taurine, betaine, and xylose) were disturbed, which could be regarded as potential biomarkers of RA. The intestinal microbiome of RA rats had changed, according to the results of 16 S rRNA gene sequencing; eight microbes (g_norank_f_Eubacterium_coprostanoligenes_group, g_Ruminococcus_torques_group, g_Dubosiell, g_Lactobacillus, g_norank_f_Desulfovibrionaceae, g_Bacteroides, g_Oscillibacter, and g_Romboutsia) occurred significantly at the genus level, and DSD significantly impacted six of them (g_Dubosiell, g_Lactobacillus, g_norank_f_Eubacterium_coprostanoligenes_group, g_Ruminococcus_torques_grou, g_Bacteroides, and g_Romboutsia). Three of them (g_norank_f_Eubacterium_ coprostanoligenes_group, g_Romboutsia, and g_Lactobacillus) were regarded as key microbiomes for DSD to treat RA, and three common metabolic pathways (taurine and hypotaurine metabolism; alanine, aspartate, and glutamate metabolism; primary bile acid biosynthesis) were discovered based on the 1H NMR metabonomics and PICRUST2 prediction of 16 S rRNA gene sequencing. Six SCFAs in feces (acetic acid, butyric acid, propionic acid, caproic acid, isobutyric acid, and valeric acid) increased significantly in RA, according to the outcomes of targeting SCFAs, while five SCFAs (acetic acid, butyric acid, propionic acid, caproic acid, and valeric acid) had decreased significantly due to DSD treatment. In conclusion, our study indicated that DSD could regulate RA's metabolic disorder by affecting intestinal microbiome and its metabolites. It also establishes a framework for future research into exploiting gut microbes therapeutic to treat RA.

[Mechanism of Danggui Sini Decoction in improving kidney injury caused by blood stasis syndrome based on metabolomics and network pharmacology].

This article analyzed the mechanism of Danggui Sini Decoction(DSD) in improving kidney injury caused by blood stasis syndrome(BSS) in rats. Firstly, 32 female SD rats were randomly divided into the following four groups: a normal group and a BSS group, both receiving an equal amount of distilled water by gavage; a normal+DSD group and a BSS+DSD group, both receiving 5.103 g·kg~(-1) DSD orally for a total of 14 days. Daily cold water bath was given to establish the BSS model, and on the 14th day, BSS rats were subcutaneously injected with 0.8 mg·kg~(-1) adrenaline. Normal rats were subjected to the water bath at 37 ℃ and injected with an equal volume of distilled water. After the experiment, 24-hour urine, serum, and kidney samples were collected for metabolomic analysis, biochemical measurements, and hematoxylin-eosin(HE) staining. The study then employed ~1H-NMR metabolomic technology to reveal the metabolic network regulated by DSD in improving BSS-induced kidney injury and used network pharmacology to preliminarily elucidate the key targets of the effectiveness of DSD. Pathological and biochemical analysis showed that DSD intervention significantly reduced inflammation and abnormal levels of blood creatinine, blood urea nitrogen, and urine protein in the kidneys. Metabolomic analysis indicated that DSD attenuated BSS-induced kidney injury primarily by regulating 10 differential metabolites and three major metabolic pathways(taurine and hypotaurine metabolism, citrate cycle, and acetaldehyde and dicarboxylic acid metabolism). Network pharmacology analysis suggested that the protective effect of DSD against BSS-induced kidney injury might be related to two key genes, ATP citrate lyase(ACLY) and nitric oxide synthase 2(NOS2), and two main metabolic pathways, i.e., arginine biosynthesis, and arginine and proline metabolism. This study, from the perspective of network regulation, provides initial insights and evidence into the mechanism of DSD in improving kidney injury induced by BSS, offering a basis for further investigation into the molecular mechanisms underlying its efficacy.

Meteorin links the bone marrow hypoxic state to hematopoietic stem/progenitor cell mobilization.

Hematopoietic stem/progenitor cells (HSPCs) are supported and regulated by niche cells in the bone marrow with an important characterization of physiological hypoxia. However, how hypoxia regulates HSPCs is still unclear. Here, we find that meteorin (Metrn) from hypoxic macrophages restrains HSPC mobilization. Hypoxia-induced factor 1α and Yin Yang 1 induce the high expression of Metrn in macrophages, and macrophage-specific Metrn knockout increases HSPC mobilization through modulating HSPC proliferation and migration. Mechanistically, Metrn interacts with its receptor 5-hydroxytryptamine receptor 2b (Htr2b) to regulate the reactive oxygen species levels in HSPCs through targeting phospholipase C signaling. The reactive oxygen species levels are reduced in HSPCs of macrophage-specific Metrn knockout mice with activated phospholipase C signaling. Targeting the Metrn/Htr2b axis could therefore be a potential strategy to improve HSPC mobilization for stem cell-based therapy.

Two ACTH analogs exert differential effects on monocytes/macrophages function regulation in ayu (Plecoglossus altivelis).

Adrenocorticotropic hormone (ACTH), a bioactive peptide of the family of melanocortins, is generated from pro-opiomelanocortin (POMC). So far, the research on the specific functions of ACTH in the immune system of teleosts is limited. We determined two complementary DNA (cDNA) sequences of POMC in ayu (Plecoglossus altivelis), termed PaPOMC-A and PaPOMC-B. PaPOMCs transcripts occurred in all examined tissues, and their expression in immune tissues changed following experimental infection with Vibrio anguillarum. PaACTH-B, but not PaACTH-A, suppressed the phagocytosis of monocytes/macrophages (MO/MФ). Two isoforms of PaACTH increased the bactericidal capacity of MO/MФ. PaACTH-A increased anti-inflammatory cytokine expression, while PaACTH-B decreased pro-inflammatory cytokine expression in MO/MФ. Compared with PaACTH-B treatment, the PaACTH-A treatment improved survival rate and reduced the bacterial load in V. anguillarum-infected ayu through interleukin (IL)-10. Our results indicate that the two PaACTH isoforms exert different effects in the host defense against bacterial infection.

Aggregation-Enhanced Sonodynamic Activity of Phthalocyanine-Artesunate Conjugates.

The clinical prospect of sonodynamic therapy (SDT) has not been fully realized due to the scarcity of efficient sonosensitizers. Herein, we designed phthalocyanine-artesunate conjugates (e.g. ZnPcT4 A), which could generate up to ca. 10-fold more reactive oxygen species (ROS) than the known sonosensitizer protoporphyrin IX. Meanwhile, an interesting and significant finding of aggregation-enhanced sonodynamic activity (AESA) was observed for the first time. ZnPcT4 A showed about 60-fold higher sonodynamic ROS generation in the aggregated form than in the disaggregated form in aqueous solutions. That could be attributed to the boosted ultrasonic cavitation of nanostructures. The level of the AESA effect depended on the aggregation ability of sonosensitizer molecules and the particle size of their aggregates. Moreover, biological studies demonstrated that ZnPcT4 A had high anticancer activities and biosafety. This study thus opens up a new avenue the development of efficient organic sonosensitizers.

Comparison of the clinical application values of PCT, hs-CRP and SAA detection in the early diagnosis of sepsis.

OBJECTIVES: To investigate the clinical application values of procalcitonin (PCT), high-sensitivity C-reactive protein (hs-CRP) and serum amyloid A (SAA) in the early diagnosis of sepsis. METHODS: In this retrospective analysis, 36 patients admitted to Liaocheng People's Hospital were selected from May 2018 to July 2019. According to infectious disease diagnostic criteria, 17 patients were confirmed to have sepsis (observation group), and 19 patients were determined to be nonseptic (control group). The levels of PCT, CRP and SAA of patients were detected on admission, and the clinical application values of PCT, CRP and SAA for sepsis were compared. RESULTS: Seventeen patients were included in the observation group, including 9 males and 8 females, with an average age of 52.18 ± 9.49 years; 19 patients were included in the control group, including 12 males and 7 females, with an average age of 51.53 ± 8.50 years. On admission, there were significant differences in white blood cell (WBC) count (t = 5.134), neutrophil count (t = 3.143), lymphocyte count (t = 2.510), PCT (t = 9.250), hs-CRP (t = 2.947) and SAA (t = 11.360) between the observation group and the control group, and the differences were statistically significant. For the comparison of clinical application values: the sensitivity of PCT, hs-CRP and SAA was 78.95%, 52.17% and 50.00%, respectively; the specificity of PCT, hs-CRP and SAA was 88.24%, 61.54% and 37.50%, respectively; the area under the ROC curve (AUC) of PCT, hs-CRP and SAA was 0.920, 0.684 and 0.870, respectively; the logistic regression coefficient of PCT, hs-CRP and SAA was -0.577, -0.028 and -0.009, respectively; and the 95% confidence interval (CI) of PCT, hs-CRP and SAA was 0.779-0.985, 0.508-0.828 and 0.716-0.958, respectively. CONCLUSION: Compared with hs-CRP and SAA, PCT had a higher clinical application value for sepsis, and PCT could be used as a reliable index for the early diagnosis of sepsis.

Impact of calcium peroxide dosage on the control of nutrients release from sediment in the anoxic landscape water.

The anoxic and reductive aquatic environment is formed easily in summer due to the global warming, which may accelerate endogenous release. In this experiment, four different dosages of calcium peroxide (CaO2) were adopted to study the control effects of nutrients release from the sediments in the simulated landscape waters. The results demonstrated that CaO2 addition could effectively improve the physicochemical properties and microbial composition in sediments, and an obvious improvement was achieved with a larger dosage. It was observed that the surface sediments of experiment groups were oxidized to form a capping barrier between the sediment and overlying water, which might cut off the pollutant diffusion in sediment. Meanwhile, CaO2 could decrease the nutrients concentration in water obviously, and the reduced effect was positively correlated with the CaO2 dosage. Compared with the nutrients release fluxes in CK (105.89 mg-TN m-2 day-1, 106.48 mg-NH4+-N m-2 day-1, 4.14 mg-TP m-2 day-1, and 4.30 mg-SRP m-2 day-1), the CaO2 dosages of 0.12 and 0.18 kg m-2 could entirely inhibit the nutrients release from sediment, and partially reduce the original pollutants in the overlying water. However, 0.18 kg m-2 CaO2 would cause a higher increase of pH value and NO2--N concentration, and bring potential risk to the aquatic ecosystem. Therefore, 0.12 kg-CaO2 m-2-sediment was selected as the optimal dosage by considering the control effect, economic cost, and potential risk comprehensively. In general, this study provided a quantitative usage method of CaO2, which is convenient and effective to prevent or control the nutrients release from sediment caused by anoxic and reductive condition in summer.

Effects and mechanisms of calcium peroxide on purification of severely eutrophic water.

In consideration of severe eutrophication of scenic water caused by pollutants released from sediments in summer, calcium peroxide (CaO2) was adopted as the slow release peroxide to improve the water quality in a field experiment of 600 m2. The effect of CaO2 on the overlying water, interstitial water, sediment, and sediment microorganisms of scenic water was studied. Results for two months indicated that the dissolved oxygen (DO) concentration of the overlying water in the test zone was 3.78 times that in the control zone; the oxidation-reduction potential (ORP) in the overlying water and sediment increased significantly (p = 0.002 and p = 0). Meanwhile, CaO2 could effectively inhibit the release of nitrogen (N) and phosphorus (P) from the sediment and could obviously reduce the concentrations of N and P in the overlying water by enhancing the microbiological action. Moreover, the average concentrations of total nitrogen and total phosphorus in the overlying water of the test zone were 46.27% and 50.51% of those in control zone, respectively, and the concentrations of N and P in the interstitial water decreased during the entire experiment. In addition, CaO2 decreased the relative abundance of anaerobic bacteria in the sediment, whereas it increased that of aerobic bacteria and promoted the appearance of the functional bacteria, such as Nitrospirae and Thermodesulfoba. In conclusion, CaO2 can improve the DO and ORP in the eutrophic water effectively and change the microbial community in the sediment to a certain extent, thereby controlling the pollutants released from the sediment and reducing the N and P concentrations in the overlying water. Thus, CaO2 can effectively realize the purification and restoration of the severely eutrophic scenic water.

Significant MRI indicators of malignancy for breast non-mass enhancement.

OBJECTIVE: To explore and evaluate new malignant predictors of breast non-mass enhancement lesions using the new BI-RADS MRI lexicon. METHODS: A dataset involving 422 consecutive women underwent breast 3.0 T MRI between January 2014 and July 2016 was assembled for this study. Each case was retrospectively reviewed by 3 radiologists. Eighty-four lesions that present non-mass enhancement in 79 patients were identified in the study. Dynamic contrast-enhanced MRI features were analyzed using univariate and multivariate analyses to identify significant indicators of malignancy. RESULTS: Of 84 non-mass enhancement lesions, 52 (61.9%) were malignant and 32 (38.1%) were benign. Segmental distribution (P = 0.015 from univariate analysis; OR = 4.739, P = 0.008 from multivariate analysis), cluster ring enhancement (P = 0.017 from univariate analysis; OR = 3.601, P = 0.032 from multivariate analysis), time-intensity curve of plateau (P = 0.002 from univariate analysis; OR = 3.525, P = 0.027 from multivariate analysis) and phase to peak (P = 0.06 from univariate analysis; OR = 6.327, P = 0.015 from multivariate analysis) were significantly different between malignant and benign lesions. CONCLUSIONS: This study demonstrated that segmental distribution, clustered ring enhancement, and short time to peak could act as new malignant predictors for breast non-mass enhancement detected on 3.0 T MRI.

[Biological effects of lipopolysaccharide, transforming growth factor-beta1 on murine bone marrow-derived dendritic cells].

AIM: To explore method of stimulating murine bone marrow-derived dendritic cells by lipopolysaccharide (LPS), transforming growth factor-beta1 (TGF-beta1) and to study their biological character. METHODS: Murine bone marrow-derived dendritic cells were cultivated with cytokine GM-CSF and IL-4 for 6 days, BMDC was stimulated by control, LPS, TGF-beta1, LPS +TGF-beta1 for 48 hours respectively.Morphological characters of BMDC were observed by a inversed microscope, surface molecules such as CD(11C), CD(80), CD(86) and MHC II were detected by flowcytometry, Interleukin-6 and interleukin-12 p70 in co-culture medium was quantified by ELISA. RESULTS: In LPS group it presented the most typical DC morphology with the highest expression of CD(80), CD(86) and MHC II, the strongest ability in mixed lymphocyte reaction, higher level of IL-6 and IL-12 p70 compared with control, TGF-beta1, LPS+TGF-beta1 (P<0.05). While in TGF-beta1 group it presented the less typical DC morphology with the lower expression of CD(80), CD(86), MHC II, weaker ability in mixed lymphocyte reaction, and lower levels of IL-6 and IL-12 p70 compared with control and LPS (P<0.05). CONCLUSION: LPS can stimulate maturation of BMDC in its late differentiation which makes it presents a more significant biological characteristics.TGF-beta1 can inhibit maturation but not differentiation of BMDC thereby can prevent its biological characteristic presentation.