Synthesis, biological evaluation and cellular localization study of fluorescent derivatives of Jiyuan Oridonin A.

Jiyuan Oridonin A (JOA) is a naturally occurring ent-kaurane diterpenoid that exhibits significant potential in the field of anti-tumor drug development. However, its detailed anti-cancer mechanism of action has not been fully understood. In order to investigate its anticancer mode of action, two series of novel fluorescent derivatives of JOA conjugated with naphthalimide dyes were synthesized, and their antitumor activity against five selected cancer cell lines (MGC-803, SW1990, PC-3, TE-1 and HGC-27) was evaluated. Compared with JOA, the anti-tumor activity of the vast majority of compounds were improved. Among them, B12 exhibited promising anti-proliferative activity against HGC-27 cells with IC50 value of 0.39 ± 0.09 µM. Fluorescence imaging studies demonstrated that probe B12 could enter HGC-27 cells in a dose-dependent and time-dependent manner and was mainly accumulated in mitochondria. Preliminary biological mechanism studies indicated that B12 was able to inhibit cell cloning and migration. Further studies suggested that B12-induced apoptosis was related to the mitochondrial pathway. Overall, our results provide new approaches to explore the molecular mechanism of the natural product JOA, which would contribute to its further development as an antitumor agent.

Different processes shape the patterns of divergence in the nuclear and chloroplast genomes of a relict tree species in East Asia.

Isolation by spatial distance (IBD), environment (IBE), and historical climatic instability (IBI) are three common processes assessed in phylogeographic and/or landscape genetic studies. However, the relative contributions of these three processes with respect to spatial genetic patterns have seldom been compared. Moreover, whether the relative contribution differs in different regions or when assessed using different genetic markers has rarely been reported. Lindera obtusiloba has been found to have two sister genetic clades of chloroplast (cpDNA) and nuclear microsatellite (nSSR), both of which show discontinuous distribution in northern and southern East Asia. In this study, we used the Mantel test and multiple matrix regression with randomization (MMRR) to determine the relative contributions of IBD, IBE, and IBI with respect to L. obtusiloba populations. Independent Mantel tests and MMRR calculations were conducted for two genetic data sets (cpDNA and nSSR) and for different regions (the overall species range, and northern and southern subregions of the range). We found a significant IBI pattern in nSSR divergence for all assessed regions, whereas no clear IBI pattern was detected with respect to cpDNA. In contrast, significant (or marginal) divergent IBD patterns were detected for cpDNA in all regions, whereas although a significant IBE was apparent with respect to the overall range, the effect was not detected in the two subregions. The differences identified in nSSR and cpDNA population divergence may be related to differences in the heredity and ploidy of the markers. Compared with the southern region, the northern region showed less significant correlation patterns, which may be related to the shorter population history and restricted population range. The findings of this study serve to illustrate that comparing between markers or regions can contribute to gaining a better understanding the population histories of different genomes or within different regions of a species' range.

PSMD4 regulates the malignancy of esophageal cancer cells by suppressing endoplasmic reticulum stress.

Proteasome 26S subunit non-ATPase 4 (PSMD4) is an important proteasome ubiquitin receptor and plays a key role in endoplasmic reticulum stress (ERS). However, the study of PSMD4 in esophageal cancer (EC) is relatively rare. Here, we found that the expression of PSMD4 was markedly enhanced in EC tissues and cell lines. The cell counting kit-8 (CCK-8) assay showed that overexpression of PSMD4 significantly enhanced Eca109 cell viability, while inhibition of PSMD4 reduced Eca109 cell viability. Knockdown of PSMD4 induced Eca109 cell apoptosis and cell cycle arrest. More importantly, knockdown of PSMD4 significantly enhanced the expression of glucose regulated protein 78, activating transcription factor 6, and p-protein kinase R-like ER kinase, indicating an enhanced ERS response in esophageal cancer cells. Compared with the control cells, brefeldin A significantly inhibited the expression of PSMD4 and increased the expression of p53-upregulated modulator of apoptosis. However, such effects were largely reversed after overexpressing PSMD4 in Eca109 cells, suggesting that silencing PSMD4 could enhance ERS-induced cell apoptosis. In summary, upregulation of PSMD4 promoted the progression of esophageal cancer mainly by reducing ERS-induced cell apoptosis.

Twenty-seven low-copy nuclear primers for Lindera obtusiloba (Lauraceae): A Tertiary relict species in East Asia.

PREMISE OF THE STUDY: To investigate a more detailed evolutionary history of Lindera obtusiloba (Lauraceae) and other Lindera species, polymorphic low-copy nuclear primers were developed. METHODS AND RESULTS: Unigenes of the L. obtusiloba transcriptome greater than 800 bp in length were randomly chosen for initial design of 168 primers. Agarose gel electrophoresis and Sanger sequencing were used to select low-copy nuclear genes. Twenty-seven primers were obtained and were used to investigate genetic diversity in 90 individuals from 24 populations. The nucleotide diversity ranged from 2.11 × 10-3 to 8.99 × 10-3, and haplotype diversity ranged from 0.57 to 0.97. These primers were also cross-amplified in L. aggregata, L. chunii, L. erythrocarpa, and L. glauca; up to 15 primers were successfully amplified in these related species. CONCLUSIONS: This methodology is effective for development of low-copy nuclear primers. The 27 primers developed here will be useful for evolutionary studies of L. obtusiloba and other Lindera species.

The Use of Recombinant 31 kDa Antigens of Trichinella spiralis for Serodiagnosis of Experimental Trichinellosis.

BACKGROUND: We have previously reported that a 31 kDa protein was screened from the excretory-secretory (ES) proteins of Tichinella spiralis muscle larvae (ML) by immunoproteomics using early infection sera, and the gene encoding a 31 kDa protein from T. spiralis was cloned and expressed in an E. coli expression system. In this study, the recombinant 31 kDa antigens were used for detection of anti-Trichinella antibodies in serum of experimentally infected mice by ELISA. METHODS: Anti-Trichinella IgG antibodies in sera of mice infected with Trichinella were assayed by ELISA with recombinant 31 kDa antigens, and its sensitivity and specificity were compared with ELISA with ES antigen. RESULTS: The sensitivity and specificity of ELISA with recombinant antigens was 96.67% (29/30) and 96.87% (62/64), compared with 100% (30/30) and 98.44% (63/64) of ELISA with ES antigens was (P>0.05). In heavily, moderately and lightly infected mice (500, 300 and 100 larvae/mouse), anti-Trichinella antibodies were firstly detected by ELISA with recombinant antigens at 8, 12 and 14 dpi, respectively; then increased rapidly with a detection rate of 100% respectively at 28, 22 and 30 dpi. While the antibodies were firstly detected by ELISA with ES antigens at 10, 8 and 10 dpi, respectively, the antibody positive rate reached 100% at 14, 12 and 22 dpi, respectively. CONCLUSION: The recombinant 31 kDa antigens of T. spirali had a good sensitivity and specificity for detecting anti-Trichinella antibodies and might be the potential diagnostic antigen for trichinellosis.

Serodiagnosis of trichinellosis by ELISA using recombinant nudix hydrolase of Trichinella spiralis.

Trichinella spiralis nudix hydrolase (TsNd) gene encoding a 46 kDa protein was expressed in Escherichia coli and the potential of recombinant TsNd protein (rTsNd) as an antigen for the serodiagnosis of trichinellosis was investigated by ELISA and compared with those of ELISA with T. spiralis muscle larval excretory-secretory (ES) antigens. The sensitivity of both ELISA was 100% (30/30), for the detection of anti-Trichinella IgG antibodies in sera of the experimentally infected mice, and the specificity of rTsNd-ELISA and ES-ELISA was 100% (54/54) and 98% (53/54), respectively (P;0.05). Serum anti-Trichinella antibodies were firstly detected by rTsNd-ELISA at 14 days post infection (dpi), then continued to increase with a detection rate of 100% at 36 dpi. The anti-Trichinella antibody levels at different times after infection were statistically different (P;0.05). The results showed that the rTsNd might be a potential candidate antigen for specific serodiagnosis of trichinellosis. But, it needs to be further evaluated with sera of the patients with trichinellosis and other helminthiasis.

(E)-2-Meth-oxy-9-(2-meth-oxy-9H-xanthen-9-yl-idene)-9H-xanthene.

The title compound, C28H20O4, was synthesized by a bimolecular Zn-HCl reduction in glacial acetic acid using the meth-oxy-substituted xanthone as a starting material. The crystal structure shows that the 2,2'-meth-oxy-bixanthenyl-idene unit is an E-type conformation anti-folded conformer. The mol-ecule lies on an inversion center. The meth-oxy group is almost coplanar with the attached benzene ring, with a C-O-C-C torsion angle of 179.38 (14)°.

[Characteristics of soil respiration components and their temperature sensitivity in a Pleioblastus amarus plantation in rainy area of West China].

To understand the characteristics of soil respiration components and their temperature sensitivity in a Pleioblastus amarus plantation in the Rainy Area of West China, a one-year periodic monitoring was conducted in a fixed plot of the plantation from February 2010 to January 2011. In the plantation, the mean annual soil respiration rate was 1.13 micromol x m(-2) x s(-1), and the soil respiration presented a clear seasonal pattern, with the maximum rate in mid-summer and the minimum rate in late winter. The contribution rates of the respiration of litter layer, root-free soil, and root to the total soil respiration of the plantation accounted for 30.9%, 20.8% and 48.3%, respectively, and the respiration of the components had a similar seasonal pattern to the total soil respiration, being related to temperature and litterfall. The annual CO2 efflux from the total soil respiration, litter layer CO2 release, root-free soil CO2 release, and root respiration was 4.27, 1.32, 0.87 and 2.08 Mg C x hm(-2) x a(-1), respectively. The total soil respiration and its components had significant positive linear correlations with litterfall, and significant positive exponential correlations with air temperature and the soil temperature at depth 10 cm. The Q10 values of total soil respiration, litter layer CO2 release, root-free soil CO2 release, and root respiration calculated based on the soil temperature were 2.90, 2.28, 3.09 and 3.19, respectively, suggesting that the temperature sensitivity of litter layer CO2 release was significantly lower than that of the total soil respiration and of its other components.

[Effects of simulated nitrogen deposition on the fine root characteristics and soil respiration in a Pleioblastus amarus plantation in rainy area of West China].

Fine root is critical in the belowground carbon (C) cycling in forest ecosystem. Aimed to understand the effects of nitrogen (N) deposition on the fine root characteristics and soil respiration in Pleioblastus amarus plantation, a two-year field experiment was conducted in the Rainy Area of West China. Four treatments with different levels of N deposition were installed, i. e., CK (0 g N x m(-2) x a(-1)), low N (5 g N x m(-2) x a(-1)), medium N (15 g N x m(-2) x a(-1)), and high N (30 g N x m(-2) x a(-1)). There were great differences in the biomass and element contents of <1 mm and 1-2 mm fine roots among the treatments. Comparing with < 1 mm fine roots, 1-2 mm fine roots had higher contents of lignin, P, and Mg, but lower contents of cellulose and Ca. Nitrogen deposition increased the biomass of < 2mm fine roots significantly, with the values being (533 +/- 89) g x m(-2) in CK, and (630 +/- 140), (632 +/- 168), and (820 +/- 161) g x m(-2) in treatments low N, medium N, and high N, respectively. The N, K, and Mg contents of <2 mm fine roots also had an obvious increase under N deposition. The annual soil respiration rate in treatments CK, low N, medium N, and high N was (5.85 +/- 0.43), (6.48 +/- 0.71), (6.84 +/- 0.57), and (7.62 +/- 0.55) t C x hm(-2) x a(-1), respectively, indicating that N deposition had obvious promotion effects on soil respiration. There were significant linear relationships between the annual soil respiration rate and the biomass and N content of <2 mm fine roots. N deposition increased the fine root biomass and promoted the root metabolism, and stimulated the rhizospheric soil respiration rate via promoting microbial activities.