Complete mitochondrial genome sequence of Aureoboletus raphanaceus (Boletales, Basidiomycota).

Aureoboletus raphanaceus is a member of boletoid mushroom, which is named after its distinctive radish smell. The mitochondrial genome and phylogenetic relationships with other boletes need to be investigated to gain a comprehensive understanding of it. In this study, we sequenced the mitochondrial genome of A. raphanaceus using next-generation sequencing technology and found that its mitochondrial genome is a circular DNA molecule measuring 42,157 bp. It consists of 15 core protein-coding genes, 27 transfer RNA genes, and two ribosomal RNA genes. The mitochondrial genome had a base composition of A (39.89%), C (11.06%), G (11.67%), and T (37.38%), with a GC content of 22.73%. A phylogenetic tree based on 22 mitochondrial genomes was constructed, which provided the first insights into the phylogenetic relationships of this species with related boletes.

A pair of new oxindole alkaloids isolated from Uncaria macrophylla.

A pair of new oxindole alkaloids, named macrophyllines C (1) and D (2), together with two known oxindole alkaloids isorhynchophylline (3) and corynoxine (4) were isolated from Uncaria macrophylla. Their structures were elucidated based on detailed spectroscopic analysis and by comparison with literature data. In addition, all the isolates were tested for their anti-HIV activities and cytotoxicities in C8166 cells and compounds 2-4 showed weak anti-HIV activities with EC50 values of 11.31 ± 3.29 µM, 18.77 ± 6.14 µM and 30.02 ± 3.73 µM, respectively.

Aspulvins A-H, Aspulvinone Analogues with SARS-CoV-2 Mpro Inhibitory and Anti-inflammatory Activities from an Endophytic Cladosporium sp.

Eight new aspulvinone analogues, aspulvins A-H (1-8) and aspulvinones D, M, O, and R (9-12), were isolated from cultures of the endophytic fungus Cladosporium sp. 7951. Detailed spectroscopic analyses were conducted to determine the structures of the new compounds. All isolates displayed different degrees of inhibitory activity against the severe acute respiratory syndrome coronavirus 2 main protease (SARS-CoV-2 Mpro) at 10 µM. Notably, compounds 9, 10, and 12 showed potential SARS-CoV-2 Mpro inhibition with IC50 values of 10.3 ± 0.6, 9.4 ± 0.6, and 7.7 ± 0.6 µM, respectively. For all compounds except 3 and 4, the anti-inflammatory activity occurred by inhibiting the release of lactate dehydrogenase (LDH) with IC50 values ranging from 0.7 to 7.4 µM. Compound 10 showed the most potent anti-inflammatory activity by inhibiting Casp-1 cleavage, IL-1ß maturation, NLRP3 inflammasome activation, and pyroptosis. The findings reveal that the aspulvinone analogues 9, 10, and 12 could be promising candidates for coronavirus disease 2019 (COVID-19) treatment as they inhibit SARS-CoV-2 infection and reduce inflammatory reactions caused by SARS-CoV-2.

Centrantheroside F, a new ionone glycoside from Centranthera grandiflora.

One new ionone glycoside, named centrantheroside F (1), together with 9 known compounds (2-10), were isolated from the roots of Centranthera grandiflora. Their structures were determined by spectroscopic data analyses and comparing with the literature data. The absolute configuration of 1 was confirmed via 2 D NMR and electronic circular dichroism (ECD). All isolated compounds were evaluated for their inhibitory activity on lipopolysaccharide (LPS)-induced nitric oxide (NO) production.

Immunomodulatory and antitumour bioactive labdane diterpenoids from Leonurus japonicus.

OBJECTIVES: Two labdane diterpenoids, leojapone B and heteronone B, were isolated from Leonurus japonicus Houtt., and their biological activity were evaluated in this study. METHODS: Human and mouse cancer cells, human peripheral blood mononuclear cells (PBMCs) and mouse macrophages (RAW264.7 cells) were used to evaluate the activity of leojapone B and heteronone B, while the in vivo effects of leojapone B were further examined in Lewis Lung Cancer tumour-bearing mice. KEY FINDINGS: In vitro studies showed that leojapone B selectively inhibited the proliferation of lung cancer cells, and both leojapone B and heteronone B inhibited the production of pro-inflammatory cytokines in activated PBMCs. In tumour-bearing mice model, lung tumours were reduced in size in mice treated with intraperitoneal injections of leojapone B at 20 and 30 mg/kg for 14 days. The population ratio of CD4+ /CD8+ T cells in mouse spleens was found to be increased, while regulatory T cells were decreased after leojapone B treatment. CONCLUSIONS: The inhibitory effects of leojapone B in mouse lung tumours were demonstrated for the first time in this study. The immunomodulatory activity of heteronone B were also demonstrated. Our findings indicated that both leojapone B and heteronone B may act as active components in L. japonicus.

Hyperbaric oxygen therapy for post-stroke depression: A systematic review and meta-analysis.

OBJECTIVES: Post-stroke depression (PSD) is common consequence of stroke. However, today the majority of PSD patients remains untreated or inadequately treated, especially in the developing countries. Herein, we performed a meta-analysis to evaluate efficacy and safety of hyperbaric oxygen (HBOT) therapy for PSD. PATIENTS AND METHODS: Seven electronic databases were comprehensively searched for randomized clinical trials (RCTs) from inception to May 2019. Outcome measures included response rate, depression severity, neurological deficit, physical disability and adverse events. RESULTS: A total of 27 RCTs involving 2250 participants were identified. Patients in HBOT group had a higher response rate than patients in control group (response rate: 69.4% vs 51.2%, odds ratio [OR] = 2.51, 95% confidence interval [CI] [1.83-3.43], P = 0.000). HBOT significantly reduced Hamilton Depression (HAMD) -17 item scores (weighted mean difference [WMD]  = -4.33, 95% CI [-4.82 to -3.84], P = 0.000), HAMD-24 item scores (WMD = -4.31, 95% CI [-5.01 to -3.62], P = 0.000), National Institute of Health Stroke Scale (NIHSS) scores (WMD = -2.77, 95% CI [-3.57 to -1.98], P = 0.000), Chinese Stroke Scale (CSS) scores (WMD = -3.75, 95% CI [-5.12 to -2.38], P = 0.000) and Modified Scandinavian Stroke Scale (MASSS) scores (WMD = -3.66, 95% CI [-6.26 to -1.06], P = 0.000). HBOT also improved Barthel Index (WMD = 10.68, 95% CI [7.98-13.37], P = 0.000). In subgroup analysis, Group A of studies with hemorrhage patients accounting for less than 20% achieved more reduction of HAMD 17-item score (WMD = -4.47, 95% CI [-5.17 to -3.77], P = 0.000) than Group B of studies with hemorrhage patients no less than 20% (WMD = -3.73, 95% CI [-4.20 to -3.26], P = 0.000). In addition, patents with HBOT along with antidepressants treatment achieve superior results than patients with antidepressants monotherapy. Patients with HBOT monotherapy achieve a slightly higher response rate than patients with antidepressants monotherapy (OR = 1.29, 95% CI [1.04-1.60], P = 0.000). Besides, HBOT group reported less adverse events (9.6%vs16.6%, P < 0.05). The most frequent side-effect of HBOT is ear pain (26 cases). CONCLUSION: Based on our pooled analysis, HBOT is effective and safe therapeutic approach for PSD. However, results should be cautiously interpreted due to a relatively poor methodological quality.

[Microwave In-situ Regeneration of Cu-Mn-Ce/ZSM Catalyst Adsorbed Toluene and Distribution of Bed Temperature].

Microwave in-situ regeneration of Cu-Mn-Ce/ZSM catalyst adsorbed toluene, distribution of fixed bed temperature, adsorption breakthrough curves of the catalyst after several regenerations and characterizations of the catalyst by BET and SEM were investigated in this study. The research indicated that regeneration effect of the catalyst adsorbed was excellent under conditions of microwave power 117 W, air flow 0.5 m3 x h(-1) and catalyst dosage of 800 g. Toluene desorbed was oxidized onto the surface of the catalyst, and the adsorption capacity of the catalyst was recovered simultaneously. Under microwave irradiation, bed temperature decreased slowly from inside to outside in horizontal level, and increased gradually from down to up in vertical level so that the highest temperature reached 250-350 degrees C at the upper sites of the bed. Sintering and agglomeration occurred on the surface of the catalyst in the course of regeneration so that the special surface area and micropore volume of the catalyst were reduced and breakthrough time was shortened, which was verified by six adsorption breakthrough curves and related characteristics of the catalyst. However, the structure of the catalyst was steady after two regenerations, and adsorption breakthrough time was kept at 70 min. The result showed that the changes of surface morphology and pore structure were positively correlated with the distribution of bed temperature.

[Catalytic oxidation of two-component VOCs and kinetic analysis ].

Catalytic oxidations of two-component volatile organic compounds (VOCs) toluene and chlorobenzene were investigated under microwave heating and tube furnace heating, respectively, and reaction kinetics were analyzed in this paper. The research indicated that competitive adsorption between toluene and chlorobenzene reduced their removal efficiencies by 3% -12% as compared to single component. 'Hot spot effect' and 'non-thermal effect' under microwave irradiation obviously enhanced conversion efficiencies of VOCs, especially, the chlorobenzene removal was increased by 31% -38%. Moreover, reaction temperature and energy consumption were both reduced under microwave heating. The dynamic calculations showed that microwave heating decreased the activation energies by 2 146 J. mol-1 and 1 450 J mol-1 for toluene and chlorobenzene, respectively, as compared with tube furnace heating. Meanwhile, microwave heating enhanced the reaction rate constants of chlorobenzene and toluene to about 35 times and 6 times of that of tube furnace heating.

Altered DNA methylation patterns of the H19 differentially methylated region and the DAZL gene promoter are associated with defective human sperm.

DNA methylation disturbance is associated with defective human sperm. However, oligozoospermia (OZ) and asthenozoospermia (AZ) usually present together, and the relationship between the single-phenotype defects in human sperm and DNA methylation is poorly understood. In this study, 20 infertile OZ patients and 20 infertile AZ patients were compared with 20 fertile normozoospermic men. Bisulfate-specific PCR was used to analyze DNA methylation of the H19-DMR and the DAZL promoter in these subjects. A similar DNA methylation pattern of the H19-DMR was detected in AZ and NZ(control), with only complete methylation and mild hypomethylation(<50% unmethylated CpGs) identified, and there was no significant difference in the occurrence of these two methylation patterns between AZ and NZ (P>0.05). However, the methylation pattern of severe hypomethylation (>50% unmethylated CpGs ) and complete unmethylation was only detected in 5 OZ patients, and the occurrence of these two methylation patterns was 8.54±10.86% and 9±6.06%, respectively. Loss of DNA methylation of the H19-DMR in the OZ patients was found to mainly occur in CTCF-binding site 6, with occurrence of 18.15±14.71%, which was much higher than that in patients with NZ (0.84±2.05%) and AZ (0.58±1.77%) (P<0.001).Additional, our data indicated the occurrence of >20% methylated clones in the DAZL promoter only in infertile patients, there was no significant difference between the AZ and OZ patients in the proportion of moderately-to-severely hypermethylated clones (p>0.05). In all cases, global sperm genome methylation analyses, using LINE1 transposon as the indicator, showed that dysregulation of DNA methylation is specifically associated with the H19-DMR and DAZL promoter. Therefore, abnormal DNA methylation status of H19-DMR, especially at the CTCF-binding site 6, is closely associated with OZ. Abnormal DNA methylation of the DAZL promoter might represent an epigenetic marker of male infertility.

[ICSI causes abnormal H3K9 methylation in the male pronuclei and growth retardation of mouse embryos].

OBJECTIVE: To evaluate the safety of intracytoplasmic sperm injection (ICSI) in the mouse model. METHODS: We simulated clinical ICSI technology and comprehensively evaluated it by parthenogenetic activation, immunofluorescence, embryo transplantation, examination of early implantation, and measurement of the crown-rump length (CRL). RESULTS: ICSI significantly reduced the ability of preimplantation embryo development of the mouse, especially after the 8-cell stage (P < 0.01). The fluorescence of H3K9 dimethylation was abnormal at the male pronuclei of the embryos derived from ICSI. Further examination of the development of the transferred ICSI embryos indicated no significant difference in the rate of early implantation at E5. 5 days as compared with normal fertilization (P = 0.6), but the percentage of "normal embryos" was decreased significantly at E9.5 days (P < 0.01). Obvious growth retardation phenotype was observed even in the normal ICSI embryos at E9.5 days. CONCLUSION: ICSI might result in growth retardation of embryos by affecting H3K9 dimethylation in the male pronuclei.

[Correlation of DNA methylation status of imprinted gene H19 ICR with oligozoospermia and asthenozoospermia].

OBJECTIVE: To study the correlation of the DNA methylation status of the imprinted gene H19 imprinting control region (ICR) with oligozoospermia and asthenozoospermia. METHODS: We eliminated chromosomal abnormality as the cause of male infertility in the subjects by karyotype analysis and detection of Y-chromosome microdeletions, and identified 18 cases of single factor-induced oligozoospermia (sperm concentration < 15 x 10(6)/ml) and 20 cases of single factor-induced asthenozoospermia (progressively motile sperm <32%) by computer-aided sperm analysis (CASA). Then we extracted genome-wide sperm DNA, treated it with bisul- fite, subjected the target gene fragments to PCR amplification and sequencing. Lastly, we analyzed the DNA methylation status of the target genes with BIQ Analyzer and processed the data using SPSS17.0. RESULTS: The DNA methylation level of the H19 ICR was increased significantly in the oligozoospermia patients ([9.19 +/- 2.45]%, P < 0.05), especially in the severe oligozoospermia males with sperm concentration < 3 x 10(6)/ml (P < 0.01), as compared with that of the 20 fertile control men ([0.30 +/- 0.06]%). However, no significant differences were found in the level ([0.30 +/- 0.07]%) and pattern of the DNA methylation of the H19 ICR (P = 0.62). Further analysis of the DNA methylation status of the CTCF-6 binding sites indicated that the DNA methylation degree was significant higher in the oligozoospermia men ([2.67 +/- 0.75]%) than in the fertile control ([0.05 +/- 0.03]%) or the asthenozoospermia group ([0.03 +/- 0.02]%), with no significant differences between the latter two (P = 0.35). CONCLUSION: The reduced DNA methylation of the H19 ICR is negatively correlated with sperm concentration but not associated with sperm motility.