Mechanistic Study of Icaritin-Induced Inactivation of Cytochrome P450 2C9.

Icaritin (ICT) is a prenylflavonoid derivative that has been approved by National Medical Products Administration for the treatment of hepatocellular carcinoma. This study aims to evaluate the potential inhibitory effect of ICT against cytochrome P450 (CYP) enzymes and to elucidate the inactivation mechanisms. Results showed that ICT inactivated CYP2C9 in a time-, concentration-, and NADPH-dependent manner with Ki = 1.896 µM, Kinact = 0.02298 minutes-1, and Kinact/Ki = 12 minutes-1 mM-1, whereas the activities of other CYP isozymes was minimally affected. Additionally, the presence of CYP2C9 competitive inhibitor, sulfaphenazole, superoxide dismutase/catalase system, and GSH all protected CYP2C9 from ICT-induced activity loss. Moreover, the activity loss was neither recovered by washing the ICT-CYP2C9 preincubation mixture nor the addition of potassium ferricyanide. These results, collectively, implied the underlying inactivation mechanism involved the covalent binding of ICT to the apoprotein and/or the prosthetic heme of CYP2C9. Furthermore, an ICT-quinone methide (QM)-derived GSH adduct was identified, and human glutathione S-transferases (GST) isozymes GSTA1-1, GSTM1-1, and GSTP1-1 were shown to be substantially involved in the detoxification of ICT-QM. Interestingly, our systematic molecular modeling work predicted that ICT-QM was covalently bound to C216, a cysteine residue located in the F-G loop downstream of substrate recognition site (SRS) 2 in CYP2C9. The sequential molecular dynamics simulation confirmed the binding to C216 induced a conformational change in the active catalytic center of CYP2C9. Lastly, the potential risks of clinical drug-drug interactions triggered by ICT as a perpetrator were extrapolated. In summary, this work confirmed that ICT was an inactivator of CYP2C9. SIGNIFICANCE STATEMENT: This study is the first to report the time-dependent inhibition of CYP2C9 by icaritin (ICT) and the intrinsic molecular mechanism behind it. Experimental data indicated that the inactivation was via irreversible covalent binding of ICT-quinone methide to CYP2C9, while molecular modeling analysis provided additional evidence by predicting C216 as the key binding site which influenced the structural confirmation of CYP2C9's catalytic center. These findings suggest the potential of drug-drug interactions when ICT is co-administered with CYP2C9 substrates clinically.

Characterization of the anti-fatigue effect of Astragali Radix from the Shanxi Hengshan area and its association with taurine metabolism based on 1H NMR metabolomic technology combined with network pharmacology / 药学学报

The objective of this work was to evaluate the anti-fatigue efficacy of Astragali Radix (AR) from the Shanxi Hengshan area and to reveal possible mechanisms by which it relieves fatigue. Efficacy differences between Guangling (GL) and Hunyuan (HY) AR preparations were compared and evaluated, and an 1H NMR metabolomic technique combined with statistical methods was used to identify the metabolites in different groups of mouse gastrocnemius muscle tissues. The differential metabolites after AR treatments were identified according to VIP and P values and the upstream targets were predicted with the help of Metscape. Cytoscape software was utilized to construct a network map of AR potential anti-fatigue targets. Key differential metabolites were identified based on shared targets and entered into the Metaboanalyst website for pathway enrichment analysis, which led to the preliminary elucidation of the molecular mechanisms. The results showed that intervention with AR can significantly improve the swimming-to-exhaustion time, increase liver glycogen, and reduce urea-nitrogen levels in mice. The difference between GL and HY ARs was relatively small, indicating that the quality of AR produced in the Hengshan area is consistent and stable. The metabolic fingerprints of mouse gastrocnemius muscle tissue extracts were composed of 34 metabolites, and the statistical results showed that 19 differential metabolites were significantly reversed after the Hengshan AR intervention. We found that the anti-fatigue effects of AR in the Shanxi Hengshan area were mainly associated with taurine and hypotaurine metabolism through regulation of GAD1, based on network pharmacological analysis. In conclusion, 1H NMR metabolomic techniques were combined with network pharmacology to compare and evaluate the quality of Hengshan ARs, and further associate the fatigue relieve with the regulation of taurine metabolism. This provides a theoretical basis for the resource utilization of Hengshan ARs and the development of anti-fatigue-related products. The animal experiments in this study followed the regulations of the Animal Ethics Committee of Shanxi University and passed the ethical review of animal experiments (Approval No. SXULL2021028).

Cardiac rehabilitation program for patients undergoing small incision aortic valve replacement / 中华胸心血管外科杂志

Objective:To study the effect of cardiac rehabilitation program on recovery of patients after small incision aortic valve replacement.Methods:600 patients who underwent small incision aortic valve replacement in our hospital from January 2015 to January 2020 were retrospectively collected and divided into Cardiac rehabilitation group(CR) and control group by propensity matching analysis. Clinical data of CR group and control group were collected 6 months and 12 months after the beginning of Cardiac rehabilitation program. The primary outcome measures were the peak oxygen uptake(VO 2 Peak) of cardiopulmonary function test and the number of patients attending cardiovascular specialty in tertiary hospitals after the rehabilitation program began. The secondary outcome measures were 6-minute Walk test(6-MWT), psychological evaluation, and assessment of cardiovascular disease risk factors. Results:After 6 months and 12 months of cardiac rehabilitation program, the VO2 peak in CR group was statistically significant compared with the control group, and the 6-MWT index after 12 months was statistically significant.The cardiovascular specialist visits in tertiary hospitals in the two groups were statistically significant. The comparison of psychological self-rating scale and cardiovascular disease risk factors between the two groups was statistically significant after propensity matching analysis.Conclusion:Cardiac rehabilitation has a positive effect on postoperative recovery after small incision aortic valve replacement, and can improve patients’ motor ability.

MiR-181a promotes the growth of chondrosarcoma cells by targeting PTEN / 中国医师杂志

Objective:To explore the effect of miR-181a on chondrosarcoma cell growth through phosphatase and tensin homolog(PTEN) and its possible regulatory mechanism.Methods:From January to December 2022, 10 fresh chondrosarcoma and 10 chondroma tissues from orthopedic patients of Hunan Provincial People′s Hospital were collected, and the expression of miR-181a in chondrosarcoma and chondroma tissues was detected using real-time fluorescence quantitative polymerase chain reaction (qRT-PCR); Chondrosarcoma cell SW1353 was cultured in vitro and transfected with miR-181a inhibitor (miR-181a inhibition group) and control (miR-NC, control group), respectively. The effects of miR-181a on the growth and proliferation of SW1353 cells were detected by cell counting kit (CCK-8) and clone formation test, respectively; The binding sites between miR-181a and PTEN were predicted through the Target Scan database, and verified using dual luciferase reporter gene experiments; The mimetic miR-181a (miR-181a group) and its control (miR-NC, control group) were transfected into chondrosarcoma cell SW1353, respectively. The adenosine triphosphate (ATP) content, glucose consumption, and lactic acid production in the cells were measured, and the effect of miR-181a on glycolysis of SW1353 cells was analyzed. Results:The expression of miR-181a in chondrosarcoma tissues was significantly higher than that in chondroma tissues ( P<0.05). The cell growth and clonogenesis ability of miR-181a inhibition group were significantly lower than those of control group (all P<0.05). It was predicted by Target Scan online website that there might be binding sites between miR-181a and PTEN, and co-transfection of wild-type PTEN and miR-181a could significantly reduce luciferase activity by double luciferase reporter assay ( P<0.05). The ATP content, glucose consumption and lactic acid production of miR-181a group were significantly higher than those of miR-NC group (all P<0.05). Conclusions:MiR-181a promotes the growth of chondrosarcoma cells through PTEN-mediated glycolysis.

First Detection of Mukawa Virus in Ixodes persulcatus and Haemaphysalis concinna in China.

Mukawa virus (MKWV), a novel tick-borne virus (TBV) of the genus Phlebovirus of family Phenuiviridae, has been firstly reported in Ixodes persulcatus in Japan. In this study, we made an epidemiological investigation in China to obtain the geographic distribution and genetic features of this virus outside Japan. We screened 1,815 adult ticks (665 I. persulcatus, 336 Dermacentor silvarum, 599 Haemaphysalis longicornis, 170 Rhipicephalus microplus, 45 Haemaphysalis concinna) and 805 wild small mammals collected from eight provinces. The positive rate of 6.77% (45/665, including 18 female and 27 male I. persulcatus) and 2.22% (1/45, 1 male H. concinna) were obtained from I. persulcatus and H. concinna in Heilongjiang province, respectively. No evidence of MKWV infection was found in other three tick species or any of the mammalian species. The virus can infect the Vero cells successfully, indicating the ability of MKWV to replicate in mammalian cells. A phylogenetic tree based on the nucleotide sequences of L, M, and S segments demonstrated that the Japanese MKWV variant, our two MKWV variants, and KURV were clustered with the members of the mosquito/sandfly-borne phleboviruses and distant from other tick-borne phenuiviruses. A phylogenetic analysis based on 895 bp partial L gene sequences (n = 46) showed that all MKWV sequences were separated into three lineages. Our results showed the presence of MKWV in I. persulcatus and H. concinna in northeast of China, highlighting the necessity of epidemiological study in wider regions. Due to the ability of MKWV to replicate in mammalian cells, the potential for zoonosis, and wide distribution of I. persulcatus and H. concinna in China, the important vectors of MKWV, further screening to more tick species, wild animals, domestic animals, and humans raises up practical significance.

Pig carcass-derived biochar caused contradictory effects on arsenic mobilization in a contaminated paddy soil under fluctuating controlled redox conditions.

Contamination of paddy soils by arsenic (As) is of great concern for human health and the environment. The impact of animal-derived biochar on As mobilization under fluctuating redox conditions in paddy soils has not been studied. Consequently, we investigated the effects of pig carcass-derived biochar (PB) on As (im)mobilization in a contaminated paddy soil under controlled redox potential (Eh) using a biogeochemical microcosm-setup. The addition of PB decreased the concentration of dissolved As at Eh = +100 and +200 mV by 38.7% and 35.4%, respectively (compared to the control), because of the co-precipitation of As with Fe-Mn oxides and the complexation between As and aromatic organic molecules. However, under reducing conditions (Eh = -300 mV), PB increased the dissolved As by 13.5% through promoting reduction and decomposition of As-bearing Fe minerals (e.g., ferrihydrite-As, Fe-humic-As). Under oxidizing conditions (Eh = +250 mV), PB increased the dissolved As by 317.6%, due to the associated increase of pH. We conclude that As mobilization in PB-treated paddy soils is highly affected by Eh. PB can be used to reduce the environmental risk of As under moderately reducing conditions, but it may increase the risk under highly reducing and oxidizing conditions in paddy soils.

Sleep-improving mechanism of Chaiqin Ningshen Granules in insomnia rats: based on hippocampal metabonomics / 中国中药杂志

With the ultra high performance liquid chromatography-quadruple-electrostatic field orbitrap high resolution mass spectrometry(UHPLC-Q Exactive Orbitrap-MS)-based metabonomics technology, this study aims to analyze the effect of Chaiqin Ningshen Granules(CNG) on endogenous metabolites in insomnia rats of liver depression syndrome and explore the sleep-improving mechanism of this prescription. Parachlorophenylalanine(PCPA, ip) and chronic stimulation were combined to induce insomnia of liver depression pattern in rats, and the effect of CNG on the macroscopic signs, hemorheology, and neurotransmitters in the hippocampus of insomnia rats of liver depression syndrome was observed. After the administration, rat hippocampus was collected for liquid chromatography-mass spectrometry(LC-MS) analysis of the metabolomics. Principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were employed for analyzing the metabolites in rat hippocampus and screening potential biomarkers. MetPA was used to yield the related metabolic pathways and metabolic networks. The results show that the drugs can significantly improve the mental state, liver depression, and blood stasis of rats, significantly increase the content of 5-hydroxytryptamine(5-HT) and gamma aminobutyric acid(GABA) in hippocampus(except low-dose CNG), and significantly reduce the content of glucose(Glu)(except low-dose CNG). Among them, estazolam and high-dose CNG had better effect than others. Metabolomics analysis yielded 27 potential biomarkers related to insomnia. MetPA analysis showed 4 metabolic pathways of estazolam in intervening insomnia and 3 metabolic pathways of high-dose CNG in intervening insomnia, involving purine metabolism, glycerophospholipid metabolism, histidine metabolism, and caffeine metabolism. CNG can alleviate insomnia by regulating endogenous differential metabolites and further related metabolic pathways. The result lays a basis for further elucidating the mechanism of CNG in improving sleep.

Meta analysis of indocyanine green fluorescence imaging guided laparoscopic liver tumor resection / 中华肝胆外科杂志

Objective:This meta-analysis was conducted to systematically evaluate the short-term efficacy and safety of indocyanine green fluorescence imaging guided laparoscopic liver tumor resection (FIGLTR).Methods:A systematic search was made for the literature on indocyanine green fluorescence image-guided laparoscopic hepatectomy in randomized, semi-randomized controlled trials and observational studies, and manually search published materials and conference papers in Chinese and English and trace references included in the literature. The retrieval period was up to September 2021. The quality of included studies was evaluated, then the meta-analysis was conducted using Review Manager 5.1 software.Results:Ten studies were included with 803 patients, including 341 in the FIGLTR group and 462 in the non-FIGLTR group. Meta results showed that: Compared to the traditional resection group, indocyanine green fluorescence imaging can significantly shorten the operative time ( MD=-22.61, 95% CI: -34.20--11.03, P<0.001), reduce intraoperative bleeding ( MD=-49.17, 95% CI: -84.99--13.36, P<0.01), shorter hospital stay ( MD=-0.89, 95% CI: -7.72--0.06, P<0.05), Improve the removal rate of R 0 edge ( OR=8.80, 95% CI: 1.96-39.44, P<0.05) and reduce the incidence of postoperative complications ( OR=0.55, 95% CI: 0.34-0.87, P<0.05) of laparoscopic liver tumor resection. There were no differences found in portal block time and transfusion rate. Conclusion:Indocyanine green fluorescence imaging technology provides a new way for safe and accurate laparoscopic resection of liver tumors.

Infection with severe fever with thrombocytopenia virus in healthy population: a cohort study in a high endemic region, China.

BACKGROUND: Severe fever with thrombocytopenia (SFTS) caused by SFTS virus (SFTSV) was a tick-borne hemorrhagic fever that posed significant threat to human health in Eastern Asia. The study was designed to measure the seroprevalence of SFTSV antibody in healthy population residing in a high endemic region. METHODS: A cohort study was performed on healthy residents in Shangcheng County in Xinyang City from April to December in 2018, where the highest SFTS incidence in China was reported. Anti-SFTSV IgG was measured by indirect enzyme-linked immunosorbent assay and neutralizing antibody (NAb) was detected by using PRNT50. The logistic regression models were performed to analyze the variables that were associated with seropositive rates. RESULTS: Totally 886 individuals were recruited. The baseline seroprevalence that was tested before the epidemic season was 11.9% (70/587) for IgG and 6.8% (40/587) for NAb, which was increased to 13.4% (47/350) and 7.7% (27/350) during the epidemic season, and further to 15.8% (80/508) and 9.8% (50/508) post epidemic. The IgG antibody-based seropositivity was significantly related to the patients aged ≥ 70 years old [adjusted odds ratio (OR) = 2.440, 95% confidence interval (CI): 1.334-4.461 compared to the group of < 50 years old, P = 0.004], recent contact with cats (adjusted OR = 2.195, 95% CI: 1.261-3.818, P = 0.005), and working in tea garden (adjusted OR = 1.698, 95% CI: 1.002-2.880, P = 0.049) by applying multivariate logistic regression model. The NAb based seropositivity was similarly related to the patients aged ≥ 70 years old (adjusted OR = 2.691, 95% CI: 1.271-5.695 compared to the group of < 50 years old, P = 0.010), and recent contact with cats (OR = 2.648, 95% CI: 1.419-4.941, P = 0.002). For a cohort of individuals continually sampled with 1-year apart, the anti-SFTSV IgG were maintained at a stable level, while the NAb level reduced. CONCLUSIONS: Subclinical infection might not provide adequate immunity to protect reinfection of SFTSV, thus highlighting the ongoing threats of SFTS in endemic regions, which called for an imperative need for vaccine development. Identification of risk factors might help to target high-risk population for public health education and vaccination in the future.

Prognostic Value of Immune-Related Multi-IncRNA Signatures Associated With Tumor Microenvironment in Esophageal Cancer.

Esophageal cancer is the eighth most common cancer and the sixth leading cause of cancer death worldwide. Hence, for a better understanding of tumor microenvironment and to seek for novel molecular targets for esophageal cancer, we performed related studies on two histopathological subtypes of esophageal cancer: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). Bioinformatic analyses were conducted based on the RNA-seq, genomic mutation, and clinical data from TCGA and GEO cohorts. We clustered patients into high-immunity and low-immunity groups through the ssGSEA results. The ESTIMATE algorithm was used to evaluate the tumor microenvironment. Patients with high immunity in both ESCC and EAC had lower tumor purity and poor survival. Subsequently, CIBERSORT was performed to learn about the detailed difference of tumor-infiltrating lymphocytes (TILs) between high- and low-immunity patients. Specific increase of M2 macrophages and decrease of activated dendric cells can be observed in ESCC and EAC, respectively. The most enriched functions and pathways of high-immunity patients were immunoglobulin complex, MHC class II protein complex, and allograft rejection according to the GO terms and KEGG. Two prognostic immune-related multi-lncRNA risk models were constructed and validated by ROC curve and PCA in ESCC and EAC. High-risk patients in both subtypes had poor survival, advanced clinical characteristics, and higher drug susceptibility except cisplatin and sorafenib. In addition, the tumor mutation burden (TMB) was positively correlated with the risk level in the ESCC and EAC and showed distinct differences between the two subtypes. In summary, we comprehensively analyzed the tumor microenvironment for two subtypes of esophageal cancer, identified two multi-lncRNA signatures predictive for the prognosis, and explored the possibility of the signatures to forecast drug susceptibility as well as TMB for the first time. The findings may serve as a conceptual basis for innovative strategy of individualized immunotherapy for esophageal cancer.

The Identification and Genetic Characterization of Parechovirus Infection Among Pediatric Patients With Wide Clinical Spectrum in Chongqing, China.

Human parechoviruses (HPeVs) are important causes of infection in children. However, without a comprehensive and persistent surveillance, the epidemiology and clinical features of HPeV infection remain ambiguous. We performed a hospital-based surveillance study among three groups of pediatric patients with acute respiratory infection (Group 1), acute diarrhea (Group 2), and hand, foot and mouth disease (Group 3) in Chongqing, China, from 2009 to 2015. Among 10,212 tested patients, 707 (6.92%) were positive for HPeV, with the positive rates differing significantly among three groups (Group 1, 3.43%; Group 2, 14.94%; Group 3, 3.55%; P < 0.001). The co-infection with other pathogens was detected in 75.2% (531/707) of HPeV-positive patients. Significant negative interaction between HPeV and Parainfluenza virus (PIV) (P = 0.046, OR = 0.59, 95% CI = 0.34-0.98) and positive interactions between HPeV and Enterovirus (EV) (P = 0.015, OR = 2.28, 95% CI = 1.23-4.73) were identified. Among 707 HPeV-positive patients, 592 (83.73%) were successfully sequenced, and 10 genotypes were identified, with HPeV1 (n = 396), HPeV4 (n = 86), and HPeV3 (n = 46) as the most frequently seen. The proportion of genotypes differed among three groups (P < 0.001), with HPeV1 and HPeV4 overrepresented in Group 2 and HPeV6 overrepresented in Group 3. The spatial patterns of HPeV genotypes disclosed more close clustering of the currently sequenced strains than those from other countries/regions, although they were indeed mixed. Three main genotypes (HPeV1, HPeV3, and HPeV4) had shown distinct seasonal peaks, highlighting a bi-annual cycle of all HpeV and two genotypes (HPeV 1 and HPeV 4) with peaks in odd-numbered years and with peaks in even-numbered years HPeV3. Significantly higher HPeV1 viral loads were associated with severe diarrhea in Group 2 (P = 0.044), while associated with HPeV single infection than HPeV-EV coinfection among HFMD patients (P = 0.001). It's concluded that HPeV infection was correlated with wide clinical spectrum in pediatric patients with a high variety of genotypes determined. Still no clinical significance can be confirmed, which warranted more molecular surveillance in the future.

Role of the miR-133a-5p/FBXO6 axis in the regulation of intervertebral disc degeneration.

OBJECTIVE: Low back pain is a leading cause of disabilities worldwide, and intervertebral disc degeneration (IVDD)-related disorders have been recognised as one of the main contributors. Nevertheless, the underlying mechanism has not yet been fully understood. The aim of this study was to investigate the role of the miR-133a-5p/FBXO6 axis in the regulation of IVDD. METHODS: RT-qPCR, WB and IHC were performed to assess the expression of FBXO6 in human IVD tissues. Nucleus pulposus (NP) cells were treated with IL-1ß to induce IVDD cellular model. Silence of FBXO6 was achieved using specific siRNAs. CCK-8 assay, flow cytometry, TUNEL assay, RT-qPCR and WB were used to evaluate the role and mechanism of FBXO6 in the process of IVDD. Online tools, GSE datasets and RT-qPCR were used to search the candidate miRNAs targeting FBXO6. The direct binding sites between FBXO6 and miR-133a-5p were further verified by a dual luciferase assay. RT-qPCR, WB and rescue experiments were conducted to identify the regulatory function of miR-133a-5p on the expression of aggrecan, collagen Ⅱ, MMP3, ADAMTS5, IL-6 and COX2. In addition, the role of the NF-κB pathway in regulating miR-133a-5p was studied using lentiviral shRNA, WB and RT-qPCR. RESULTS: Results showed that FBXO6 mainly expressed in the NP tissue of IVD and the expression of FBXO6 decreased with the process of IVDD as well as under IL-1ß stimulation. The silence of FBXO6 led to the decreased expression of aggrecan and collagen Ⅱ and the increased expression of MMP3, ADAMTS5, IL-6 and COX2, which further induced the degeneration of NP cells. The bioinformatic analysis showed that miR-133a-5p was the candidate miRNA targeting FBXO6. miR-133a-5p was upregulated in IVDD tissues and significantly inhibited the expression of FBXO6. The inhibition of miR-133a-5p ameliorated the acceleration of IVDD induced by the silence of FBXO6 in vitro. Moreover, it was demonstrated that IL-1ß regulated the expression of the miR-133a-5p/FBXO6 axis via the NF-κB pathway in NP cells. CONCLUSION: miR-133a-5p was upregulated by IL-1ß to aggravate intervertebral disc degeneration via sponging FBXO6. Inhibiting miR-133a-5p expression or rescuing FBXO6 expression may be promising strategies for the treatment of IVDD. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: This study suggests that the miR-133a-5p/FBXO6 axis could regulate NP cells proliferation, apoptosis, synthesis and degradation of extracellular matrix, which provides a promising therapeutic target and strategy for the treatment of IVDD.

LRRK2 is a candidate prognostic biomarker for clear cell renal cell carcinoma.

BACKGROUND: Clear cell renal cell carcinoma (ccRCC), derived from renal tubular epithelial cells, is the most common malignant tumor of the kidney. The study of key genes related to the pathogenesis of ccRCC has become important for gene target therapy. METHODS: Bioinformatics analysis of The Cancer Genome Atlas (TCGA), the NCBI Gene Expression Omnibus (GEO) database, USUC Xena database, cBioPortal for Cancer Genomics, and MethSurv were performed to examine the aberrant genetic pattern and prognostic significance of leucine-rich repeat kinase 2 (LRRK2) expression and its relationship to clinical parameters. Immunohistochemistry and Western blot were performed to verify LRRK2 expression. The regulation of ccRCC tumor cell lines proliferation by LRRK2 was examined by CCK8 assay. RESULTS: Bioinformatics analysis showed that LRRK2 expression was up-regulated and largely correlated with DNA methylation in ccRCC. The up-regulation of LRRK2 was confirmed in ccRCC tissue immunohistochemically and by protein analysis. The level of expression was related to gender, pathological grade, stage, and metastatic status of ccRCC patients. Meanwhile, Kaplan-Meier analysis showed that high expression of LRRK2 correlates to a better prognosis; knockdown of LRRK2 expression attenuated the proliferation ability of ccRCC tumor cell lines; protein-protein interaction network analysis showed that LRRK2 interacts with HIF1A and EGFR. CONCLUSION: We found that LRRK2 may play an important role in the tumorigenesis and progression of ccRCC. Our findings provided a potential predictor and therapeutic target in ccRCC.

Immobilization of cadmium and lead using phosphorus-rich animal-derived and iron-modified plant-derived biochars under dynamic redox conditions in a paddy soil.

Functionalized biochar has gained extensive interests as a sustainable amendment for an effective remediation of paddy soils contaminated with heavy metals (HMs). We examined the efficiency of pig carcass-derived biochar (P-rich biochar, total P = 8.3%) and pristine (raw biochar, total Fe = 0.76%) and Fe-modified (Fe-rich biochar, total Fe = 5.5%) green waste-derived biochars for the immobilization of cadmium (Cd) and lead (Pb) in a paddy soil under pre-defined redox conditions (Eh, from -400 to +300 mV). Average concentrations (µg L-1) of dissolved Cd increased under reducing conditions up to 10.9 in the control soil, and decreased under oxidizing conditions to below the detection limit (LDL = 2.7) in the raw and Fe-rich biochar treated soils. Application of the raw biochar decreased the concentrations of dissolved Cd by 43-59% under Eh ≤ -100 mV, compared to the non-treated control, which was more effective than the Fe-rich biochar (31-59%) and the P-rich biochar (8-19%). The immobilization of Cd under low Eh might be due to its precipitation with sulfide (S2-), whereas its immobilization under high Eh might be due to the associated increase of pH. Concentrations (µg L-1) of Pb ranged from 29.4 to 198.2 under reducing conditions, and decreased to LDL (12.5) under oxidizing conditions. The P-rich biochar was more effective in immobilizing Pb than the raw and Fe-rich biochars, particularly under Eh ≤ 0 mV (55-82%), which might be due to the retention of Pb by phosphates. The raw and Fe-rich biochars immobilized Pb under low Eh (≤ -300 mV), but both biochars, particularly the Fe-rich biochar mobilized Pb under Eh higher than -200 mV, especially at +100 mV, due to the decrease of pH at this point (pH = 6.0 to 6.5). These results improved our understanding of using P-rich and Fe-rich functionalized biochars for the immobilization of Cd and Pb in a paddy soil under stepwise redox changes. The amendment of P-rich pig carcass-derived biochar to paddy soils could be a promising approach for mitigating the risk of Pb for human health and the environment. The raw and Fe-rich green waste-derived biochars can be used for immobilizing Cd and mitigating its risk in paddy soils under both reducing and oxidizing conditions.

Exosomes from human umbilical cord mesenchymal stem cells attenuate the inflammation of severe steroid-resistant asthma by reshaping macrophage polarization.

BACKGROUND: Severe, steroid-resistant asthma (SSRA) is a serious clinical problem in asthma management. Affected patients have severe clinical symptoms, worsened quality of life, and do not respond to steroid, a mainstay steroid treatment of asthma. Thus, effective therapies are urgently needed. Exosomes derived from mesenchymal stem cell (MSC-Exo) has become attractive candidates for the lung inflammatory diseases through its immunomodulatory effects. In this study, we explored the therapeutic effects of MSC-Exo in SSRA and identified the therapeutic mechanism of MSC-Exo. METHOD: Exosomes from human umbilical cord mesenchymal stem cell (hUCMSC) were isolated and characterized by transmission electron microscopy, nanoparticle tracking analysis and flow cytometry analysis. Effects of MSC-Exo on airway hyper responsiveness (AHR), inflammation, histopathology, and macrophage polarization in SSRA in mice were evaluated. Systematic depletion of macrophages determined the role of macrophages in the therapeutic effect of SSRA in mice. LPS-stimulated RAW 264.7 cell model was constructed to determine the underlying mechanism of MSC-Exo on macrophage polarization. qRT-PCR, Western blotting, immunofluorescence, and flow cytometry were performed to evaluate the expression of M1 or M2 markers. Tandem mass tags (TMT)-labeled quantitative proteomics were applied to explore the central protein during the regulation effect of MSC-Exo on macrophage polarization. Knockdown and overexpression of TRAF1 were used to further clarify the role of the central protein on macrophage polarization. RESULT: We successfully isolated and characterized exosomes from hUCMSCs. We verified that the intratracheal administration of MSC-Exo reversed AHR, histopathology changes, and inflammation in SSRA mice. Systematic depletion of macrophages weakened the therapeutic effect of MSC-Exo. We found that MSC-Exo treatment inhibited M1 polarization and promoted M2 polarization in LPS-stimulated RAW 264.7 cells. Subsequently, tumor necrosis factor receptor-associated factor 1 (TRAF1) was determined as the central protein which may be closely related to the regulation of macrophage polarization from TMT-labeled quantitative proteomics analysis. Knockdown and overexpression of TRAF1 demonstrated that the effect of MSC-Exo treatment on macrophage polarization, NF-κB and PI3K/AKT signaling was dependent on TRAF1. CONCLUSION: MSC-Exo can ameliorate SSRA by moderating inflammation, which is achieved by reshaping macrophage polarization via inhibition of TRAF1.

Three-dimensional multi-gene expression maps reveal cell fate changes associated with laterality reversal of zebrafish habenula.

The conserved bilateral habenular nuclei (HA) in vertebrate diencephalon develop into compartmentalized structures containing neurons derived from different cell lineages. Despite extensive studies demonstrated that zebrafish larval HA display distinct left-right (L-R) asymmetry in gene expression and connectivity, the spatial gene expression domains were mainly obtained from two-dimensional (2D) snapshots of colorimetric RNA in situ hybridization staining which could not properly reflect different HA neuronal lineages constructed in three-dimension (3D). Combing the tyramide-based fluorescent mRNA in situ hybridization, confocal microscopy and customized imaging processing procedures, we have created spatial distribution maps of four genes for 4-day-old zebrafish and in sibling fish whose L-R asymmetry was spontaneously reversed. 3D volumetric analyses showed that ratios of cpd2, lov, ron, and nrp1a expression in L-R reversed HA were reversed according to the parapineal positions. However, the quantitative changes of gene expression in reversed larval brains do not mirror the gene expression level in the obverse larval brains. There were a total 87.78% increase in lov+ nrp1a+ and a total 12.45% decrease in lov+ ron+ double-positive neurons when the L-R asymmetry of HA was reversed. Thus, our volumetric analyses of the 3D maps indicate that changes of HA neuronal cell fates are associated with the reversal of HA laterality. These changes likely account for the behavior changes associated with HA laterality alterations.

Pristine and iron-engineered animal- and plant-derived biochars enhanced bacterial abundance and immobilized arsenic and lead in a contaminated soil.

In this study, typical animal- and plant-derived biochars derived from pig carcass (PB) and green waste (GWB), and their iron-engineered products (Fe-PB and Fe-GWB) were added at the dose of 3% (w/w) to an acidic (pH = 5.8) soil, and incubated to test their efficacy in improving soil quality and immobilizing arsenic (As = 141.3 mg kg-1) and lead (Pb = 736.2 mg kg-1). Soil properties, microbial activities, and the geochemical fractions and potential availabilities of As and Pb were determined in the non-treated (control) and biochar-treated soil. Modification of PB (pH = 10.6) and GWB (pH = 9.3) with Fe caused a decrease in their pH to 4.4 and 3.4, respectively. The application of PB and GWB significantly increased soil pH, while Fe-PB and Fe-GWB decreased soil pH, as compared to the control. Application of Fe-GWB and Fe-PB decreased the NH4H2PO4-extractable As by 32.8 and 35.9%, which was more effective than addition of GWB and PB. However, PB and GWB were more effective than Fe-PB and Fe-GWB in Pb immobilization. Compared to the control, the DTPA-extractable Pb decreased by 20.6 and 21.7%, respectively, following PB and GWB application. Both biochars, particularly PB significantly increased the 16S rRNA bacterial gene copy numbers, indicating that biochar amendments enhanced the bacterial abundance, implying an alleviation of As and Pb bio-toxicity to soil bacteria. The results demonstrated that pristine pig carcass and green waste biochars were more effective in immobilizing Pb, while their Fe-engineered biochars were more effective in As immobilization in co-contaminated soils.

Efficacy and safety of CalliSpheres microsphere versus conventional transcatheter arterial chemoembolization in treatment of hepatocellular carcinoma: A Meta-analysis / 临床肝胆病杂志

ObjectiveTo investigate the efficacy and safety of CalliSpheres microsphere-transcatheter arterial chemoembolization (CSM-TACE) versus conventional transcatheter arterial chemoembolization (cTACE) in the treatment of hepatocellular carcinoma (HCC) through a meta-analysis. MethodsPubMed, Web of Science, Cochrane Library, CNKI, Wanfang Data, and VIP were searched for all Chinese and English articles on the application of CSM-TACE and cTACE in HCC published up to the end of October, 2020. After quality assessment was performed for the articles included, RevMan 5.3 software provided by Cochrane Library was used for analysis. ResultsA total of 15 studies were included, with 1535 patients in total. This meta-analysis showed that compared with the patients receiving cTACE, the patients receiving CSM-TACE had significantly higher 1-year overall survival rate (odds ratio ［OR］=2.26, 95% confidence interval ［CI］: 1.63-3.13, P＜0.000 01), 2-year overall survival rate (OR=1.73, 95%CI: 1.20-2.50, P=0.003), and 2-year progression-free survival rate (OR=1.60, 95%CI: 1.05-2.43, P=0.03). In terms of safety, compared with the patients receiving cTACE, the patients receiving CSM-TACE had significantly lower incidence rates of postoperative vomiting (OR=0.65, 95%CI: 0.46-0.92, P=0.01), bone marrow suppression (OR=0.17, 95%CI: 0.05-0.54, P=0.003), and neutropenia (OR=0.18, 95%CI: 0.07-045, P=0.000 3), while there were no significant differences between the two groups of patients in postoperative pyrexia, abdominal pain, and ascites (all P＞0.05). ConclusionCSM-TACE has significant advantages in improving 1- and 2-year overall survival rates and 2-year progression-free survival rates and can significantly reduce the incidence rates of postoperative vomiting, bone marrow suppression, and neutropenia. Therefore, CSM-TACE is a safe and effective treatment method.

The complete chloroplast genome sequence of Citrus maxima (Burm.) Merr. 'Guanximiyou'.

Citrus maxima (Burm.) Merr. 'Guanximiyou' is a major citrus tree largely cultivated in China. A previous study has reported the complete chloroplast genome of C. maxima, but there may be some differences between wild species and cultivating variety. In this study, the complete chloroplast genome sequence of 'Guanximiyou' pummelo was characterized using BGISEQ-500 sequencing. The chloroplast genome was 160,186 bp in length and separated into four distinct regions such as large single-copy region (87,939 bp), small single-copy region (18,395 bp), and a pair of inverted repeat regions (26,926 bp). The genome contained a total of 109 genes including 79 protein-coding genes, 29 tRNA genes, and 4 rRNA genes. Phylogenetic maximum-likelihood analysis revealed that 'Guanximiyou' pummelo was clustered with other Rutaceae species with high bootstrap values.