[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.

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.

[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.