Blockade of the P2Y2 Receptor Attenuates Alcoholic Liver Inflammation by Targeting the EGFR-ERK1/2 Signaling Pathway.

Purpose: It is well known that inflammation plays a key role in complex pathological progressions of alcohol-associated liver disease (ALD). To date, effective therapy for ALD is lacking. P2Y2 receptor (P2Y2R), a G protein-coupled P2Y purinergic receptor, represents a novel pharmacological target in many inflammations. Methods: The alcohol-associated liver injury and inflammation mouse model was established. The effect of P2Y2R on alcohol-induced liver injury and inflammation was evaluated using quantitative real-time PCR, Western blot and immunohistochemical assay. An alcohol-stimulated (100 mmol/L, for 24 h) AML-12 cell model was established. Different agonists, antagonists and P2Y2R siRNA were used to explore the possible mechanisms of P2Y2R. Results: In vivo, results showed that the hepatoprotective effect of P2Y2R blockade by significantly suppressed liver structural abnormalities and lipid infiltration, and decreased levels of ALT/AST and TNF-α/IL-1ß in the high dosage group of suramin (20 mg/kg) compared to control diet (CD)-fed mice. At the same time, we found that alcohol feeding promoted the phosphorylation of EGFR and ERK1/2, both of which were effectively inhibited by suramin (20 mg/kg). In vitro, suramin or P2Y2R silencing effectively inhibited the phosphorylation of EGFR and ERK1/2, similar to the down-regulated effects of their corresponding inhibitors (EGFR inhibitor AG1478 and ERK1/2 inhibitor U0126) accompanied by reduced levels of TNF-α and IL-1ß compared to alcohol-induced AML-12 cell. In addition, we found that silencing P2Y2R attenuated the apoptosis of hepatocyte. Conclusion: Our findings suggest that P2Y2R regulates alcoholic liver inflammation by targeting the EGFR-ERK1/2 signaling pathway and plays an important role in hepatocyte apoptosis, which may provide new ideas for the development of methods to treat ALD.

[Effects of Earthworms/Biochar on Bacterial Diversity and Community in As-contaminated Red Soil].

Cerium-manganese modified biochar (MBC) combined with earthworms (Eisenia foetida) can immobilize arsenic (As) in red soils. In this study, high-throughput sequencing technology was used to explore the combined effects of MBC and E. foetida on bacterial diversity and community structure in As-contaminated red soils. The results showed that the single earthworm treatment had the highest diversity index, whereas the diversity index decreased in the single biochar or MBC treatment, indicating that earthworms can boost the growth of bacteria in the soil, and the addition of biochar/MBC all decreased the bacterial diversity of soils. When biochar/MBC was combined with earthworms, the diversity index increased to some degree. In terms of bacterial community structure, the relative abundance of Proteobacteria and Bacteroidetes increased significantly in each treatment, especially for MBC-earthworm treated soil, in which the relative abundance was increased by 17.08% and 329.47% for Proteobacteria and Bacteroidetes, respectively, compared to that in the control (CK). Otherwise, those abundances were decreased by 19.18% and 48.76%, respectively, for Acidobacteria and Chloroflexi. Correlation analysis results showed that the soil water-soluble As (WSAs) was negatively correlated with the relative abundances of Proteobacteria and Bacteroides (P<0.05) but was positively correlated with Acidobacteria and Chloroflexi (P<0.05), which indicated that with the decrease in WSAs in soils, the bacteria of Proteobacteria and Bacteroides reproduced rapidly, whereas the Acidobacteria and Chloroflexi were inhibited. Moreover, different treatments induced selective changes in the bacterial community, in which earthworms significantly promoted the proliferation of γ-Proteobacteria, Flavobacteriales, Aeromonadales, and Variovorax and earthworms improved the immobilization effect of As by promoting the growth of these bacteria.

Slow magnetic relaxation in structurally similar mononuclear 8-coordinate Fe(II) and Fe(III) compounds.

A pair of structurally-similar and stable 8-coordinate high-spin Fe(ii) and Fe(iii) compounds have been obtained. Both compounds exhibit field-induced slow magnetic relaxation behaviour. The Fe(iii) compound represents the first example of 8-coordinate Fe(iii) single-molecule magnets (SMM).

Tunable Luminescent Properties of Tricyanoosmium Nitrido Complexes Bearing a Chelating O

We have recently reported a strongly luminescent osmium(VI) nitrido complex [OsVI(N)(NO2-L)(CN)3]- [HNO2-L = 2-(2-hydroxy-5-nitrophenyl)benzoxazole]. The excited state of this complex readily activates the strong C-H bonds of alkanes and arenes (Commun. Chem. 2019, 2, 40). In this work, we attempted to tune the excited-state properties of this complex by introducing various substituents on the bidentate L ligand. The series of nitrido complexes were characterized by IR, UV/vis, 1H NMR, and electrospray ionization mass spectrometry. The molecular structures of five of the nitrido compounds have been determined by X-ray crystallography. The photophysical and electrochemical properties of these complexes have been investigated. The luminescence of these nitrido complexes in the solid state, in a CH2Cl2 solution, and in a CH2Cl2 solid matrix at 77 K glassy medium clearly shows that these emissions are due to 3LML'CT [L ligand to Os≡N] phosphorescence. The presence of strongly electron-withdrawing substituents in these complexes enhances the LML'CT emission. Our result demonstrates that the excited-state properties of this novel class of luminescent osmium(VI) nitrido complexes can be fine-tuned by introducing various substituents on the bidentate L ligand.

Field-induced slow magnetic relaxation in low-spin S = 1/2 mononuclear osmium(v) complexes.

Photochemical reactions of (PPh4)[OsVI(N)(L)(CN)3] (NO2-OsN) with piperidine and pyrrolidine afforded two osmium(v) hydrazido compounds, (PPh4)[OsV(L)(CN)3(NNC5H10)] ([PPh4]1) and (PPh4)[OsV(L)(CN)3(NNC4H8)] ([PPh4]2), respectively. Their structures consist of isolated, mononuclear distorted octahedral osmium anions that are well-separated from each other by PPh4+. Their low spin S = 1/2 and L = 1 ground state was confirmed by magnetometry and DFT calculations. Interestingly, both compounds exhibit slow magnetic relaxation under a bias dc-field. These osmium(v) complexes are potentially useful building-blocks for the construction of molecule-based architectures with interesting magnetic properties. In contrast, the structurally related (PPh4)[OsIII(L)(CN)3(NH3)] ([PPh4]3), which also has a low-spin S = 1/2 ground state but with a different electronic configuration (5d5), does not exhibit slow magnetic relaxation, due to the absence of any orbital moment (L = 0). Furthermore, the structurally different osmium(v) hydrazido compound reported by Meyer, [OsV(tpy)(Cl)2(NNC5H10)](PF6) (4[PF6]), also does not exhibit slow magnetic relaxation due possibly to a change in magnetic anisotropy from axial for [PPh4]1 and [PPh4]2 to planar.

Synthesis and reactivity of an osmium(iii) aminoguanidine complex.

The biological activities of aminoguanidine (GNH2) and its derivatives have been extensively studied due to their properties as radical scavengers and antioxidants. Some of their biological activities may result from their binding to various metals present in biological systems. However, the reactivity of coordinated aminoguanidines has not been investigated. We report herein the synthesis, structure and reactivity of a novel osmium(iii) complex bearing the parent aminoguanidine, mer-[Os{NHC(NH2)(NHNH2)}(L)(CN)3]- (OsGNH2, HL = 2-(2-hydroxyphenyl)benzoxazole). The antioxidant properties of OsGNH2 have been investigated by reactions with various oxidants, including O2, H2O2, m-chloroperbenzoic acid (m-CPBA) and Ce(iv). Various osmium products are produced, which depend on the type of oxidant used. OsGNH2 is readily oxidized by O2 or H2O2 under ambient conditions to afford an osmium(iii) formamidine complex, [OsIII(NH2C[double bond, length as m-dash]NH)(L)(CN)3]- (OsFA, FA = formamidine). With m-CPBA, the nitrosyl complex, mer-[Os(NO)(L)(CN)3]- (OsNO), is formed instead. On the other hand, the nitrido complex mer-[Os(N)(L)(CN)3]- (OsN) is produced when the one-electron oxidant (NH4)2[CeIV(NO3)6] (Ce(iv)) is employed. The molecular structures of OsGNH2 and OsFA have been determined by X-ray crystallography. The oxidation of OsGNH2 to OsFA by O2 or H2O2 is proposed to go through initial dehydrogenation to give a diazoamidine intermediate. In the oxidation by m-CPBA and Ce(iv), it is proposed that the initially formed OsFA is further oxidized to OsNO and OsN, respectively, via osmium(iii) hydrogen cyanamido and osmium(iv) cyanoimido intermediates.

Purinergic P2X7 receptor blockade mitigates alcohol-induced steatohepatitis and intestinal injury by regulating MEK1/2-ERK1/2 signaling and egr-1 activity.

The P2X7 receptor is an ATP-binding cation channel involved in a broad range of inflammatory diseases. However, little is known about the potential role of P2X7R in alcohol-induced steatohepatitis and intestinal injury. In our study, C57BL/6 mice were intraperitoneally injected with P2X7R antagonists Brilliant Blue G and A438079 from the 4th day to the 10th day during the induction of chronic plus binge alcohol feeding model. Our results showed that alcohol feeding induced significant steatohepatitis and liver injury, which were mitigated by P2X7R blockade as evidenced by decreased serum levels of ALT, AST, T-CHO and TG, reduced lipid accumulation, and less inflammation. The increased intestinal inflammatory cytokines production and the prominent intestinal barrier disruption caused by alcohol were also modulated by P2X7R antagonism. Interestingly, alcohol feeding increased the relative abundance of phylum Bacteroidetes while decreased the number of phylum Verrucomicrobia and genus Akkermansia in the cecal content, which were reversed by P2X7R antagonist. Importantly, the improvement of intestinal barrier function and the restoration of partial taxonomic alterations in the gut microbiota might contribute to protect the liver from gut microbiota dysbiosis-induced second hit. Furthermore, P2X7R blockade inhibited MEK1/2-ERK1/2 phosphorylation and egr-1 expression in both liver and intestine from alcohol-fed mice. Collectively, P2X7R blockade mitigates alcohol-induced steatohepatitis and intestinal injury by inhibiting MEK1/2-ERK1/2 signaling and egr-1 expression. These studies strongly suggest that P2X7R blockade may be a promising therapeutic approach for treating alcoholic liver disease.

Polynuclear Cu(i) and Ag(i) phosphine complexes containing multi-dentate polytopic ligands: syntheses, crystal structures and photoluminescence properties.

A series of polynuclear metal complexes, [Cu2(L1)(PPh3)4](ClO4)2 (1), [Cu3(L2)(PPh3)6](ClO4) (2), [Cu3(L3)(PPh3)6] (3), [Ag2(L1)(PPh3)4](BF4)2 (4), [Ag4(L2)2(PPh3)6] (5) and [Ag3(L3)(PPh3)5] (6), have been obtained from the reactions of the highly conjugated bridging ligands 2,3-bis(2-pyridyl)pyrazine (L1), 2,3-bis(2-tetrazoyl)pyrazine (H2L2) and 2,3-bis(2-tetrazoyl)imidazole (H3L3) with [Cu(MeCN)4]ClO4 and AgBF4, respectively. Their crystal structures have been determined by X-ray crystallography and their photophysical properties have been investigated in detail. Complexes 1 and 3 show photoluminescence in CH2Cl2 solution, while all the complexes exhibit obvious luminescence in the solid state; detailed photophysical studies and density functional theory calculations of these complexes have revealed that their lowest energy absorptions and emissions are predominantly derived from either metal-to-ligand charge-transfer (MLCT) or intraligand (IL) excited states.

Responses in reproductive organs, steroid hormones and CYP450 enzymes in female Mongolian gerbil (Meriones unguiculatus) over time after quinestrol treatment.

The aim of this study was to assess the effects and reversibility of the synthetic estrogen compound, quinestrol, on the reproductive organs, steroid hormones, and drug-metabolizing enzymes CYP3A4 and CYP1A2 in liver and kidney over time after two quinestrol treatments in female Mongolian gerbils (Meriones unguiculatus). Female gerbils were treated with 4mg/kg quinestrol (9 gerbils/group, 3 treated group) (1 control group, 0mg/kg) for 3days and treated again after 25days. Animals were killed for collection of samples at 5, 10 and 15days after the second treatment ending. Two interval quinestrol treatments significantly increased uterine weight, with trend of increase over time, but no change could be detected in ovarian weights. Quinestrol treatment increased progesterone and estradiol levels, both with trend of decline over time. Quinestrol increased liver and kidney weights and total enzyme content of CYP3A4 and CYP1A2, with trend of decline over time. On the basis of reversible changes of detoxification enzymes or organs, interval quinestrol treatment effectively and reversibly influenced the reproductive hormone and organ to some extent.

Call divergence in three sympatric Rattus species.

To reduce errors in species recognition and the probability of interbreeding that lowers fitness, individuals within sympatric zones shift the signals to differentiate from those of other species. In the present study, the differences of the acoustic features of ultrasounds (courtship calls during heterosexual encounters) and audible calls (distress calls during tail-clamp stress) are compared among three sympatric Rattus species (Rattus andamanensis, R. norvegicus, and R. losea). Results showed that the three species have significantly different call parameters, including call duration, peak frequency, bandwidth, pitch, goodness of pitch, frequency modulation, and Wiener entropy. This study provides quantitative evidence for character displacement in the acoustic signals of closely related sympatric Rattus species. Results indicate that the divergence of acoustic signal has arrived at the quite meticulous degree of evolution. Therefore, the acoustic signal trait is likely involved in the evolution of species diversity in rodents.

Effects of quinestrol on the vocal behavior of mice during courtship interactions.

Vocalizations are a crucial part of courtship and mating in a wide variety of species. Mating behavior, including courtship calls, is modulated by sex steroid hormones. Male mice produce courtship ultrasonic vocalizations to attract females during heterosexual encounters. However, rare is the knowledge on whether vocal behavior of mice changes under sterilant treatment which will affect gonadal hormone levels. In the present study, we treat male mice with quinestrol, which interferes with the release of the gonadotropin-releasing hormone (GnRH) and has a significant anti-fertility effect in rodents. We compared the differences in the syllable structures (including peak intensity, peak frequency, duration, and bandwidth), total number of calls, and harmonic syllable proportions between quinestrol treated and control male mice. Male mice treated with quinestrol produced more courtship calls and more harmonic syllables than control mice, whereas the parameters of call syllables showed no significant change between the two groups. The results indicate that normal male vocal behavior during sexual interactions could be retained or even reinforced after quinestrol treatment. In addition, female mice approached male mice treated with quinestrol more than control mice, suggesting that the treated male mice were more attractive to the female mice than the controls. Thus, competitive reproductive interference is enhanced. Further, findings provided behavior mechanism in vocal context of the fertility control in mice.

Assessment of effects of mifepristone administration to lactating mice on the development and fertility of their progeny.

AIM: Mifepristone, a synthetic steroid compound that induces abortion, was recently found to potentially pass into human milk. This study aimed to determine the effects of mifepristone administration to lactating mice on the development and reproduction of their progeny. METHODS: Lactating mice were gavage fed with mifepristone (8 mg/kg) daily for 4 days either from days 1-4 or from days 7-10 of lactation; controls received only peanut oil. Growth, mortality rate, organ weight to bodyweight ratio, sex hormone at 20, 40 and 60 days, fertility of these F1 progeny and litter size, sex ratio and mortality rate of the second generation were recorded. RESULTS: No significant differences were observed in the average bodyweight, mortality rate of the female or male pups, and organ coefficient of uterus and ovaries of females in adulthood in comparison with the controls. However, the organ coefficient of testis at day 20 and 40 and testosterone concentration at day 60 were increased in male pups. Moreover, the reproductive capacity of the F1 pups was unaffected by 4 days exposure to mifepristone via their mother's milk: time to birth of F2 pups, litter size, sex ratio and mortality rate were similar to control F1 pups. CONCLUSION: The study showed that treating lactating mice with 8 mg/kg mifepristone influenced only the organ coefficient of testis at day 40 and the testosterone concentration in male pups at day 60, however, it did not affect the development and fertility of female and male pups.

[Expression of miR-16 in patients with T lymphoblastic lymphoma/acute lymphoblastic leukemia].

This study was purposed to investigate the expression of miR-16 in T lymphoblastic lymphoma/acute lymphoblastic leukemia (T-LBL/ALL) and its relation with target therapy and prognosis. The CD3, cCD3, CD10, CD20, CD34, CD43, CD99, TdT, PAX-5, BCL-2 and Ki67 in paraffin samples from 38 cases of T-LBL/ALL were detected by immunohistochemical labeling; the miR-16 expression level was detected by real-time RT-PCR. Fifteen cases of reactive hyperplasia of lymph nodes were selected as control. The results indicated that among 38 cases of T-LBL/ALL the positive rate of TdT was highest (94.7%), the positive rate of CD34 was lowest (22.1%), the PAX-5 and CD20 were found to be negative. The Ki67 expression level in 39.5% cases exceeded 80%. As compared with reactive hyperplasia of lymph node, the miR-16 expression in T-LBL/ALL was up-regulated, ant its expression level was 4.87-fold of reactive hyperplasia of lymph node (P < 0.05). The overall survival rate in group of miR-16 high expression decreased (P < 0.05). The prognosis of T-LBL/ALL patients with BCL-2 positive expression was better than that of patients with BCL-2 negative expression (P < 0.05). The miR-16 expression correlated with BCL-2 protein (r = 0.51, P < 0.05). It is concluded that the overall survival rate in miR-16 high expression group is higher than that in miR-16 low expression group, suggesting possible relation of miR-16 with prognosis. Moreover, the prognosis in BCL-2 positive expression group is better than that in negative expression group, which may be a factor influencing prognosis.

[Study on HPLC fingerprint of jinzhen oral solution].

The HPLC fingerprint determination method of Jinzhen oral solution was established to provide a new method for quality control of Jinzhen oral solution. RP-HPLC was used for phenomenex Luna C18 (4.6 mm x 250 mm, 5 µm) chromatographic column, with 0.1% H3 PO4 water solution and acetonitrile as the mobile phase for gradient elution. The detection wavelength was 280 nm. HPLC fingerprint of Jinzhen oral solution was established to identify 17 common peaks in Jinzhen oral solution. The similarity of fingerprints of 10 batches of finished products was more than 0. 90. The established HPLC fingerprint has a better precision, reproducibility and stability, and can be applied in quality control of Jinzhen oral solution.

[Expression of blood Th17 and CD4(+) CD25(+) Treg cells in patients with aplastic anemia].

This study was aimed to investigate the expression of blood Th17 and CD4(+) CD25(+) regulatory T cells (Treg) in the patients with aplastic anemia (AA). Forty-five patients with AA were enrolled into this study, and were divided into mild aplastic anemia (MAA) group (n = 25) and severe aplastic anemia group (SAA) (n = 20), blood cell count was recorded. 15 healthy donors were enrolled as control. Proportions of blood Th17 and CD4(+) CD25(+) Treg cells were determined by flow cytometry. The serum levels of IL-17, IFN-γ and TNF-α, as well as their concentrations in culture supernatant of phytohemagglutinin (PHA) -stimulated peripheral blood mononuclear cells, were measured by ELISA. The results showed that the proportions of blood Th17 cells and concentration of blood serum IL-17 and IFN-γ increased in patients with SAA, compared with MAA and normal controls, but CD4(+) CD25(+) Foxp3(+) Treg cells obviously decreased in patients with SAA. The concentrations of IL-17 and IFN-γ significantly increased in culture supernatant of SAA group. Hemoglobin level in the patients with AA negatively correlated with the population of Th17 cells and serum IL-17 level, whereas positively correlated with the expression of CD4(+) CD25(+)Treg cells. It is concluded that the increased response of Th17 cells and deficiency of CD4(+) CD25(+) Treg cells present in severe aplastic anemia. The severity of anemia may be related with the imbalance between Th17 and CD4(+) CD25(+)Treg cell response.