[Efficacy of Zhibai Dihuang Decoction on energy metabolism of spermatogenic cells in Ureaplasma urealyticum-infected rats: A study based on AMPK and PPARα signaling pathways].

OBJECTIVE: To study the effect of the serum containing Zhibai Dihuang Decoction (ZDD) on the energy metabolism of spermatogenic cells infected with Ureaplasma urealyticum (UU) in rats and its action mechanism. METHODS: Healthy male SD rats were randomly divided into six groups, normal control, UU-infection (UUI) model control, doxycycline, and low-, medium- and high-dose ZDD-containing serum. After successful establishment of the UUI model in vivo in the latter five groups, the rats in the normal control group were treated with simple serum and those in the latter five with respective agents. Then primary spermatogenic cells were harvested from the rats for examination of the apoptosis of spermatogenic cells, contents of lactate dehydrogenase (LDH) and adenosine triphosphate (ATP), glucose disposal rate (GDR) and expressions of AMPK and PARα proteins in the spermatogenic cells, and other related parameters. RESULTS: The apoptosis rate of the spermatogenic cells was dramatically increased in the UUI model controls compared with that in the normal controls (ï¼»49.24 ± 0.86ï¼½% vs ï¼»10.09 ± 0.52ï¼½%, P < 0.01), but significantly decreased in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»11.21 ± 1.02ï¼½%, ï¼»30.64 ± 0.99ï¼½%, ï¼»35.54 ± 1.17ï¼½% and ï¼»42.95 ± 1.31ï¼½%) in comparison with that in the UUI model control group (P < 0.01).The content of LDH in the spermatogenic cells was also remarkably increased in the UUI model controls compared with that in the normal controls (ï¼»201.12 ± 2.88ï¼½ vs ï¼»60.72 ± 1.83ï¼½) mU/ml, P < 0.01), but significantly decreased in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»90.66 ± 1.61ï¼½, ï¼»94.74 ± 1.20ï¼½, ï¼»101.24 ± 2.03ï¼½ and ï¼»111.04 ± 3.35ï¼½ mU/ml) in comparison with that in the UUI model control group (P < 0.01). The GDR in the spermatogenic cells was markedly reduced in the UUI model controls compared with that in the normal controls (ï¼»49.42 ± 1.70ï¼½% vs ï¼»99.86 ± 1.26ï¼½%, P < 0.01), but significantly elevated in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»86.90 ± 2.03ï¼½%, ï¼»84.14 ± 1.21ï¼½%, ï¼»80.30 ± 1.37ï¼½% and ï¼»75.18 ± 1.76ï¼½% in comparison with that in the UUI model control group (P < 0.01). The content of ATP was also dramatically decreased in the UUI model controls compared with that in the normal controls (ï¼»19.76 ± 1.46ï¼½ vs ï¼»58.94 ± 1.95ï¼½ µmol/L, P < 0.01), but significantly increased in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»48.34 ± 1.34ï¼½, ï¼»42.82 ± 1.30ï¼½, ï¼»38.70 ± 2.03ï¼½ and ï¼»34.78 ± 0.82ï¼½ µmol/L) in comparison with that in the UUI model control group (P < 0.01). The mitochondrial membrane potential was remarkably elevated in the UUI model controls compared with that in the normal controls (ï¼»8.53 ± 0.71ï¼½% vs ï¼»2.43 ± 0.25ï¼½%, P < 0.01), but markedly reduced in the doxycycline and low-, medium- and high-dose ZDD groups (ï¼»3.92 ± 0.36ï¼½%, ï¼»4.43 ± 0.27ï¼½%, ï¼»4.65 ± 0.22ï¼½% and ï¼»4.88 ± 0.10ï¼½% in comparison with that in the UUI model control group (P < 0.01). The phosphorylation levels of AMPK and PPARα proteins were significantly up-regulated in the UUI model controls compared with that in the normal controls (P < 0.01), but down-regulated in a dose-dependent manner in the ZDD groups. CONCLUSIONS: Zhibai Dihuang Decoction can significantly improve the damage to the mitochondrial structure and inhibit UU infection-induced apoptosis of spermatogenic cells and secretion of LDH by increasing the ATP content and GDR and regulating the phosphorylation of AMPK and PARα signaling pathway.

[Accumulation and Translocation of Cd in Brassica rapa Under the Influence of Selenium].

Cadmium (Cd) is a highly toxic heavy metal. Brassica rapa (pak choi) is a vastly common vegetable, which readily accumulates Cd. Given the current conditions of Cd contamination in domestic soil, it is important to reduce Cd accumulation in the edible part of pak choi. Research has shown that selenium (Se) can regulate Cd uptake by plants. Cd accumulation (three cultivars) and Cd uptake kinetics in pak choi were investigated under hydroponic conditions. Results showed that the three levels of selenite significantly reduced Cd content in the Hangzhouyoudonger shoot by 50%, while the levels in Suzhouqinggen and Shanghaiqing shoots were not significantly decreased with elevated levels of selenite. Selenite reduces the Cd translocation factors, and higher levels had more obvious effects; 50 µmol·L-1 of selenite significantly decreased the factors by 50% in Hangzhouyoudonger and Suzhouqinggen shoots. Selenite also increased iron (Fe) and manganese (Mn) contents in pak choi, especially in the Hangzhouyoudonger shoot, where 50 µmol·L-1 increased the Fe content by approximately 50%. In the uptake kinetics of Cd, both selenite and selenate significantly increased Cd uptake rates and Vmax by over 100%. Therefore, Se could reduce Cd accumulation in pak choi. This also depended on the tested cultivar. Therefore, reduction effects of Se on the Cd content mainly stemmed from the alteration of Cd translocation in pak choi instead of the uptake competition between Cd and Se.

[Toxicity of Chromium to Root Growth of Barley as Affected by Chromium Speciation and Soil Properties].

Tri-and hexavalent chromium have different chemical properties, and their levels of toxicity to plants are different. However, there is no limit set by the soil environmental quality risk control standard for Cr(Ⅲ) or Cr(Ⅵ). Therefore, studying the ecological toxicity of Cr has important implications for protecting the environment. Based on the dynamics of the Cr(Ⅲ) and Cr(Ⅵ) levels in soil solution collected from eight soils, the toxicity thresholds of the two Cr forms to barley roots were investigated through model calculation and correlation analysis under different soil properties. The results showed that both Cr forms and the soil properties had significant effects on the root length of barley. The effective concentrations of Cr(Ⅲ) added to the soils that led to 10% inhibition (EC10), 50% inhibition (EC50), and no-observed-effect concentrations (NOEC) were significantly higher than those of Cr(Ⅵ). The EC50 of Cr(Ⅲ) ranged from 298.8 to 2014.1 mg·kg-1 (6.7-fold variation); the EC50 of Cr(Ⅵ) ranged from 8.0 to 126.6 mg·kg-1 (15.8-fold variation). Under the same soil conditions, the EC50 of Cr(Ⅲ) was 2.8 to 101.7 times higher than that of Cr(Ⅵ), suggesting the higher phytotoxicity of Cr(Ⅵ) than Cr(Ⅲ). Correlation analysis showed that the pH and soil organic matter were the main factors that influenced the Cr toxicity thresholds, as indicated by the root length of barley. The concentration of chromium in the soil solution was below the detection limit of the TAS-990 when Cr(Ⅲ) was applied at 1280 mg·kg-1 (or less) to soils, whereas for Cr(Ⅵ), the level was 40 mg·kg-1 (or less). Cr(Ⅲ) adsorption to the soil was significantly stronger than that of Cr(Ⅵ). The toxicity of Cr(Ⅵ) was significantly higher than that of Cr(Ⅲ), which was also influenced by soil properties.

Bioinformatics Analysis Identifies Protein Tyrosine Kinase 7 (PTK7) as a Potential Prognostic and Therapeutic Biomarker in Stages I to IV Hepatocellular Carcinoma.

BACKGROUND Worldwide, hepatocellular carcinoma (HCC) accounts for 80-90% of all cases of primary liver cancer, and is one of the ten most common malignancies. This study used bioinformatics analysis to identify genes associated with patient outcome in stages I-IV HCC and the gene pathways that distinguished between normal liver and liver cells and HCC and human HCC cell lines. MATERIAL AND METHODS Target genes were defined as those that had marketed drugs or drugs under development targeting a specific gene and acquired from the Clarivate Analytics Integrity Database. Differential expression gene analysis, co-expression network analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, survival analysis and receiver operating characteristic (ROC) curve analysis were used to explore the similarities and differences in gene expression profiles, functional associations, and survival in stage I-IV HCC. Normal liver cells (HL-7702) and HCC cell lines (HepaRG, HepG2, SK-Hep1, and Huh7) were studied using Western blot and quantitative reverse transcription PCR (RT-qPCR). RESULTS Hierarchical gene clustering identified target genes that distinguished between HCC and normal liver tissue. For stages I-IV HCC, there were seven commonly upregulated target genes EPHB1, LTK, NTRK2, PTK7, TBK1, TIE1, and TLR3, which were mainly involved in immune and signaling transduction pathways. PTK7 was highly expressed in stage I-IV HCC and was an independent prognostic marker for reduced overall survival (OS). CONCLUSIONS Bioinformatics analysis, combined with patient survival analysis, identified PTK7 gene expression as a potential therapeutic target and prognostic biomarker for all stages of HCC.

Selenium Uptake and Biotransformation in Brassica rapa Supplied with Selenite and Selenate: A Hydroponic Work with HPLC Speciation and RNA-Sequencing.

Vegetables are an ideal source of human Se intake; it is important to understand selenium (Se) speciation in plants due to the distinct biological functions of selenocompounds. In this hydroponic study, the accumulation and assimilation of selenite and selenate in pak choi (Brassica rapa), a vastly consumed vegetable, were investigated at 1-168 h with HPLC speciation and RNA-sequencing. The results showed that the Se content in shoots and Se translocation factors with selenate addition were at least 10.81 and 11.62 times, respectively, higher than those with selenite addition. Selenite and selenate up-regulated the expression of SULT1;1 and PHT1;2 in roots by over 240% and 400%, respectively. Selenite addition always led to higher proportions of seleno-amino acids, while SeO42- was dominant under selenate addition (>49% of all Se species in shoots). However, in roots, SeO42- proportions declined substantially by 51% with a significant increase of selenomethionine proportions (63%) from 1 to 168 h. Moreover, with enhanced transcript of methionine gamma-lyase (60% of up-regulation compared to the control) plus high levels of methylselenium in shoots (approximately 70% of all Se species), almost 40% of Se was lost during the exposure under the selenite treatment. This work provides evidence that pak choi can rapidly transform selenite to methylselenium, and it is promising to use the plant for Se biofortification.

Difference between selenite and selenate in selenium transformation and the regulation of cadmium accumulation in Brassica chinensis.

Se can regulate Cd accumulation and translocation in plants; however, such effects can be controversial because of the differences in plant species and Se species. In this study, pak choi was cultured under hydroponic conditions, and the effects of selenite and selenate on Cd accumulation were investigated in the edible parts of this vegetable. The results showed gradual improvements in the effects of the two Se species on the Cd content in pak choi shoots at the four assessed growing stages. Selenite did not lead to significant changes in Cd accumulation in the shoots until day 40, when it significantly reduced the accumulation by 34%. Selenate was always found to increase the Cd content in the shoots, and the differences on days 19 and 40 were 16% and 45%, respectively, compared with those of the Cd (only) treatment. Accordingly, selenate invariably enhanced Cd translocation from the roots to the shoots, whereas selenite insignificantly reduced the translocation only on day 40. Generally, selenomethionine (SeMet) accounted for much larger proportions in selenite-treated plants, while SeO42- was the dominant Se species in selenate-treated plants. However, under both Se treatments, the SeMet proportion increased substantially from day 19 to day 40 when that of SeO42- exhibited a drastic decrease; therefore, the relative proportion of seleno-amino acids to SeO42- may be the key factor for the regulation of Cd accumulation in pak choi via treatment with selenite and selenate at the different growing stages.

[Differentiation of HL-60 cells induced by realgar nano-particles].

OBJECTIVE: To investigate the proliferation inhibition and the differentiation effects of realgar (As4S4) nano-particles on human acute myeloid leukemia cell line HL-60. METHOD: Cell viability was determined by MTT and PI-stained cell cycle assays. The realgar induced morphological changes on cells were examined after Wright-Giemsa staining. The cell differentiation was evaluated with NBT and specific cell surface antigen (CD11b and CD14) expression assays. RESULT: HL-60 cells exhibited obvious morphological features of differentiation after the realgar treatment. A 24 h incubation of the cells with 0.25-1.0 micromol x L(-1) realgar caused a great increase in NBT reduction ability. The expressions of CD11b and CD14 were augmented in cells treated with 0.50 micromol x L(-1) realgar for 48 h, and cell cycles were arrested in G1 phase. CONCLUSION: Low dose realgar induces differentiation in human acute myeloid leukemia cell line HL-60.

Induction of human promyelocytic leukemia HL-60 cell differentiation into monocytes by arsenic sulphide: involvement of serine/threonine protein phosphatases.

The clinical efficacy of arsenic sulfide (As(4)S(4)), also known as realgar, in the treatment of leukemia in China is prompting people to explore the underlying mechanism. We examined the realgar-induced differentiation in human promyelocytic leukemia cell line HL-60. Cells exhibited proliferation inhibition when treated with 0.10-1.5 microM of realgar, and underwent monocytic differentiation as indicated by morphological changes, NBT reduction assay, and cytofluorometric analyses of the cell surface antigens, CD11b and CD14. Accompanying the differentiation, the activity of serine/threonine protein phosphatase type 1 (PP1) and type 2A (PP2A) were enhanced, whereas the activity of PP2B remained virtually the same compared to the control. When cells were treated with realgar in the presence of an inhibitor of PP1 and 2A or an inhibitor of PP2B, the differentiation of the cells was partially suppressed as revealed by NBT reduction assay and the expression of CD14. Our data demonstrate that realgar induces monocytic differentiation in HL-60 cells and that some serine/threonine protein phosphatases may be involved in the process.

[Speciation analysis of arsenic in traditional Chinese medicine].

Some of traditional Chinese medicine (TCM) contain arsenide, such as realgar. The total amount of arsenic in the TCM exceeds the limits according to related regulations. But the roles of arsenic in TCM or its side-effects depend on its species existed in those therapies, not the total amount of arsenic. Therefore, in recent years, the analysis of arsenic in TCM focuses on the species of arsenic. The present paper summarized some methods and techniques in the speciation analysis of arsenic in TCM, in order that optimal methods can be chosen and the roles of arsenic could be evaluated properly.