Fortunella venosa (Champ. ex Benth.) C. C. Huang and F. hindsii (Champ. ex Benth.) Swingle as Independent Species: Evidence From Morphology and Molecular Systematics and Taxonomic Revision of Fortunella (Rutaceae).

Recently, the systematic status of Fortunella Swingle and its taxonomy has attracted much attention. Flora of China incorporates Fortunella into Citrus Linn. and treats all species of the traditional Fortunella as one species, namely Citrus japonica (Thunb.) Swingle. Furthermore, F. venosa (Champ. ex Benth.) C. C. Huang and F. hindsii (Champ. ex Benth.) Swingle are currently considered as synonyms of C. japonica. In this paper, morphological, palynological, and phylogenetic analyses were used to systematically explore the taxonomic status of traditional Fortunella. The key morphological features that differed among the Fortunella species were the leaf and the petiole hence could be key in its taxonomic classification of the species. Additionally, pollen morphological analysis based on the pollen size, germination grooves, polar, and equatorial axes also supported the separation of the species. The results of the phylogenetic analysis showed that each of the three species clustered separately, hence strongly supporting the conclusion of independent species. In addition, the phylogenetic analysis showed that the two genera clustered closely together hence our results support the incorporation of Fortunella into Citrus. Based on the above, this article has revised the classification of the traditional Fortunella and determined that this genus has three species, namely; F. venosa, F. hindsii, and F. japonica. F. venosa and F. hindsii are placed in the Citrus as separate species, and their species names still use the previous specific epithet. The revised scientific names of the new combinations of F. venosa and F. hindsii are as follows: Citrus venosa (Champ. ex Benth.) K. M. Liu, X. Z. Cai, and G. W. Hu, comb. nov. and Citrus hindsii (Champ. ex Benth.) K. M. Liu, G. W. Hu, and X. Z. Cai, comb. nov. F. venosa is the original species of Fortunella, F. venosa and F. hindsii are both listed as the second-class key protected wild plants in China. Therefore, the establishment of the taxonomic status of F. venosa and F. hindsii not only deepens our understanding, importance, and the complexity of the systematic classification of Fortunella, but is also significant for global biodiversity conservation, genetic resources for breeding purposes, and population genetics.

Effects of an Alternating Magnetic Field/Ag Multi-Alloying Combined Solidification Process on Cu⁻14Fe Alloy.

An alternating magnetic field (AMF)/Ag multi-alloying combined process was applied to the solidification of Cu⁻14Fe alloy to study its effects on the microstructure and properties of the resulting samples. The applied AMF and Ag multi-alloying had positive effects on the refinement of the primary Fe phase and precipitation of Fe solute atoms, respectively. These results indicated that the combined AMF/Ag multi-alloying process was effective to improve the distribution of the primary Fe phase and reduce the Fe content of the Cu matrix, which increased the conductivity of the alloy. The application of the combined AMF/Ag multi-alloying process to the solidification of Cu⁻Fe alloy provided samples with improved comprehensive properties compared with those of samples solidified using a single process (AMF or Ag multi-alloying).

[Combined effects of typical natural estrogens with MCF-7 proliferation assay].

OBJECTIVE: To study 17ß-estradiol (E2), ethinylestradiol (EE2), estriol (E3), estrone (E1) on MCF-7 proliferation effects, and compare the effects of independent action (IA) model with concentration addition (CA) model in assessing the combined effects of estrogen. METHODS: The combinations of E2 + EE2, E2 + E3 and E2 + E1 were chosen and the cellular proliferation effects were examined by MTT assay. RESULTS: The maximum proliferation effects at dose of 10⁻9 mol/L was 325.48% for E2, 330.34% for EE2, 255.22% for E3, and 199.61% for E1. In the E2 + EE2, E2 + E3, E2 + E1 groups, the results of IA model analysis were very close to the experimental results. The IA model tend to overestimated the experimental results, while the CA model often underestimated the experimental results. In the EC (E2, 30) + C (EE2, 70) group, the results exceed the maximum estrogen effects of E2, while in other groups, the results were lower. CONCLUSIONS: The estrogenic effects of the four tested substances from high to low efficiency were that: EE2 > E2 > E3 > E1. The effect of IA model in predicting the combined effects of binary mixture was better than CA model. A small proportion of binary mixture showed synergy.

[Effects of lead exposure on protein kinase C and calmodulin expression in hippocampus of baby-rats].

OBJECTIVE: To observe the effect of chronic lead contaminant on protein expression of protein kinase (PKC) and calmodulin (CaM) in hippocampus of baby-rats. METHODS: The Wistar pregnant rats were randomly divided into 3 groups fed with distilled water and lead-contained water (0.2% and 1.0% lead acetate) respectively. The lead exposure period ranged from the 0 day of pregnancy to the offspring weaned. Then the baby-rats were fed with lead water the same as their mothers. Pups were killed at postnatal day 8 and 50 respectively. Atomic absorption spectrometry was used to determine lead content of rats' brain. Western-blotting was used to observe protein expression of PKC and CaM in hippocampus of baby-rats. RESULTS: The brain lead content of test groups was much higher than that of the control group in the same growth period (P < 0.01). The content of brain lead in rats of postnatal day 50 was significantly higher than that of rats of postnatal day 8 (P < 0.01). Compared with control group, PKC and CaM protein expressions of chronic lead exposure baby-rats in the hippocampus were down trend (P < 0.05). CONCLUSION: The decrease of PKC and CaM protein expression level in hippocampus might be one of the molecular mechanisms of lead induced impairment of learning and memory.

Influence of ethanol-diesel blended fuels on diesel exhaust emissions and mutagenic and genotoxic activities of particulate extracts.

This study was aimed at evaluating the influence of ethanol addition on diesel exhaust emissions and the toxicity of particulate extracts. The experiments were conducted on a heavy-duty diesel engine and five fuels were used, namely: E0 (base diesel fuel), E5 (5%), E10 (10%), E15 (15%) and E20 (20%), respectively. The regulated emissions (THC, CO, NOx, PM) and polycyclic aromatic hydrocarbon (PAH) emissions were measured, and Ames test and Comet assay, respectively, were used to investigate the mutagenicity and genotoxicity of particulate extracts. From the point of exhaust emissions, the introduction of ethanol to diesel fuel could result in higher brake specific THC (BSTHC) and CO (BSCO) emissions and lower smoke emissions, while the effects on the brake specific NOx (BSNOx) and particulate matters (BSPM) were not obvious. The PAH emissions showed an increasing trend with a growth of ethanol content in the ethanol-diesel blends. As to the biotoxicity, E20 always had the highest brake specific revertants (BSR) in both TA98 and TA100 with or without metabolizing enzymes (S9), while the lowest BSR were found in E5 except that of TA98-S9. DNA damage data showed a lower genotoxic potency of E10 and E15 as a whole.

Protective effects of hepatocellular canalicular conjugate export pump (Mrp2) on sodium arsenite-induced hepatic dysfunction in rats.

Arsenic is a double-edged sword to human health. The excretion of various organic anions into bile is mediated by an adenosine triphosphate-dependent conjugate export pump, which has been identified as the canalicular isoform of the multidrug resistance protein 2 (Mrp2). It has been proved that Mrp2 can transport arsenite in vitro, but its effects in vivo are not clear. The aim of this study was to investigate whether Mrp2 plays a role in exportation of arsenic in vivo and its protective effects on liver function. Mrp2 protein level in rat liver was determined by Western blot analysis. Total arsenic concentrations in whole blood and bile were measured using hydride generation atomic absorption spectrometry. Alanine aminotransferase (ALT) activity, aspartate aminotransferase activity (AST), glutathione peroxidase (GSH-PX) activity, malon dialdehyde (MDA) and total bilirubin were measured by biochemical assays. The morphological changes were observed by electron microscopy. Total arsenic levels in blood and bile of arsenite-treated rats were significantly higher than those of control rats (P<0.05) at all three different time points. The overexpression of Mrp2 was 36.61%, 32.36% and 12.73% at 2, 4 and 6 weeks, respectively (percentage of controls, P<0.05), which was significantly higher than controls. A positive correlation between Mrp2 expression level and total arsenic concentration in bile indicated that Mrp2 accelerated the transport of arsenic. Electron microscopy showed that microvilli of bile canaliculi became swollen and sparse. ALT and AST activities in serum were markedly raised at 6 weeks. MDA level in serum increased (P<0.05) and GSH-PX activity in serum decreased except for 2 weeks. Damage of liver function became worse following decreased expression of Mrp2. In conclusion, overexpression of Mrp2 may explain increased biliary excretion of arsenic and it may protect liver function.

[Study on the genotoxicity of exhausts of diesel engine with ethanol-diesel blending fuel].

OBJECTIVE: To study the genotoxicity of components of diesel engine exhausts with ethanol-diesel blending fuel. To provide scientific arguments to find more economical and less polluted fuels. METHODS: Ames test, comet assay and GC-MS technique were used to test the genotoxicity and 16 kinds of PAHs on diesel engine exhausts with different proportions of ethanol (E0, E5, E10, E20). RESULTS: Both Ames test and comet assay were positive. It shows that diesel engine exhausts can lead to mutation and DNA damage, especially in pure diesel oil. But the content of 16 kinds of PAHs and DNA damage level decreased in exhausts of E5. With the increase of ethanol proportion in diesel oil, the content of 16 kinds of PAHs and DNA damage level increased. CONCLUSION: Compared with pure diesel oil and high proportion of ethanol fuel, E5 can reduce the genotoxicity and the brake specific exhausts of PAHs.