[Influence of climate change and human activities on grassland phenology in Anhui Province]. / 安徽省气候变化和人类活动对草地物候的影响.

To explore the influence of climate change and human activities on grassland phenology in Anhui Pro-vince, and quantify the contribution rate of climate change and human activities to phenology, we extracted the phenology of grassland, including the start of growing season (SOS) and the end of growing season (EOS), based on the normalized difference vegetation index (NDVI) dataset of Anhui Province from 2003 to 2020. The temporal and spatial characteristics and future evolution trends of phenological changes were analyzed using slope trend ana-lysis, Mann-Kendall non-parametric test, and Hurst index. We further conducted correlation analysis and residual analysis based on the datasets of mean annual temperature and mean annual precipitation to explore the responses of phenology to climate change and human activities, and quantify their contribution rate. The results showed that SOS and EOS showed an advancing trend with a rate of 0.8 and 0.7 days per year from 2003 to 2020. SOS in the sou-thern part of the study area was significantly earlier than in the central and northern regions, while EOS gradually advanced from south to north. Both SOS and EOS in the future showed an advancing trend. SOS was negatively correlated with annual average temperature, while positively correlated with annual precipitation. EOS was negatively correlated with both annual average temperature and annual precipitation. The proportion of the area where SOS was advanced driven by both climate change and human activities was 56.9%, and the value was 48.3% for EOS. Human activities were the main driving factor for phenology, and climate change was the secondary driving factor. The relative contributions of human activities and climate change to SOS were 66.4% and 33.6%, and to EOS were 61.2% and 38.8%, respectively. Human activities had stronger impact on SOS and EOS than climate change, resulting in earlier phenology.

Murine hematopoietic stem cell dormancy controlled by induction of a novel short form of PSF1 by histone deacetylase inhibitors.

Hematopoietic stem cells (HSCs) can survive long-term in a state of dormancy. Little is known about how histone deacetylase inhibitors (HDACi) affect HSC kinetics. Here, we use trichostatin A (TSA), a histone deacetylase inhibitor, to enforce histone acetylation and show that this suppresses cell cycle entry by dormant HSCs. Previously, we found that haploinsufficiency of PSF1, a DNA replication factor, led to attenuation of the bone marrow (BM) HSC pool size and lack of acute proliferation after 5-FU ablation. Because PSF1 protein is present in CD34(+) transiently amplifying HSCs but not in CD34(-) long-term reconstituting-HSCs which are resting in a dormant state, we analyzed the relationship between dormancy and PSF1 expression, and how a histone deacetylase inhibitor affects this. We found that CD34(+) HSCs produce long functional PSF1 (PSF1a) but CD34(-) HSCs produce a shorter possibly non-functional PSF1 (PSF1b, c, dominantly PSF1c). Using PSF1a-overexpressing NIH-3T3 cells in which the endogenous PSF1 promoter is suppressed, we found that TSA treatment promotes production of the shorter form of PSF1 possibly by inducing recruitment of E2F family factors upstream of the PSF1 transcription start site. Our data document one mechanism by which histone deacetylase inhibitors affect the dormancy of HSCs by regulating the DNA replication factor PSF1.

DNA damage enhanced by the attenuation of SLD5 delays cell cycle restoration in normal cells but not in cancer cells.

SLD5 is a member of the GINS complex composed of PSF1, PSF2, PSF3 and SLD5, playing a critical role in the formation of the DNA replication fork with CDC45 in yeast. Previously, we had isolated a PSF1 orthologue from a murine hematopoietic stem cell DNA library and were then able to identify orthologues of all the other GINS members by the yeast two hybrid approach using PSF1 as the bait. These GINS orthologues may also function in DNA replication in mammalian cells because they form tetrameric complexes as observed in yeast, and gene deletion mutants of both PSF1 and SLD5 result in a lack of epiblast proliferation and early embryonic lethality. However, we found that PSF1 is also involved in chromosomal segregation in M phase, consistent with recent suggestions that homologues of genes associated with DNA replication in lower organisms also regulate cellular events other than DNA replication in mammalian cells. Here we analyzed the function of SLD5 other than DNA replication and found that it is active in DNA damage and repair. Attenuation of SLD5 expression results in marked DNA damage in both normal cells and cancer cells, suggesting that it protects against DNA damage. Attenuation of SLD5 delays the DNA repair response and cell cycle restoration in normal cells but not in cancer cells. These findings suggest that SLD5 might represent a therapeutic target molecule acting at the level of tumor stromal cells rather than the cancerous cells themselves, because development of the tumor microenvironment could be delayed or disrupted by the suppression of its expression in the normal cell types within the tumor.

Requirement of SLD5 for early embryogenesis.

SLD5 forms a GINS complex with PSF1, PSF2 and PSF3, which is essential for the initiation of DNA replication in lower eukaryotes. Although these components are conserved in mammals, their biological function is unclear. We show here that targeted disruption of SLD5 in mice causes a defect in cell proliferation in the inner cell mass, resulting in embryonic lethality at the peri-implantation stage, indicating that SLD5 is essential for embryogenesis. Moreover, this phenotype of SLD5 mutant mice is quite similar compared with that of PSF1 mutant mice. We have previously reported that haploinsufficiency of PSF1 resulted in failure of acute proliferation of bone marrow hematopoietic stem cells (HSCs) during reconstitution of bone marrow ablated by 5-FU treatment. Since SLD5 was highly expressed in bone marrow, we investigated its involvement in bone marrow reconstitution after bone marrow ablation as observed in PSF1 heterozygous mutant mice. However, heterozygous deletion of the SLD5 gene was found not to significantly affect bone marrow reconstitution. On the other hand, abundant SLD5 expression was observed in human cancer cell lines and heterozygous deletion of the gene attenuated tumor progression in a murine model of spontaneous gastric cancer. These indicated that requirement and dependency of SLD5 for cell proliferation is different in different cell types.

[Application of zebrafish microarray on the toxicity mechanism study of bisphenol A].

The toxicity mechanism of bisphenol A (BPA) to zebrafish (Danio rerio) was studied in the molecular level,by the method of zebrafish microarray and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Zebrafish embryos were exposed to 0.5, 1.5 and 4.5 mg/L BPA for 8 days since fertilization. The results from microarray and validated by qRT-PCR showed that, 50 specific genes were up-or down-regulated, and dose-responses for them were significant (p < 0.05). This study demonstrates the genotoxicity of BPA. Moreover,according to their function and pathway analysis,it could support the mechanisms for the morphological toxicity and metabolize turbulence observed in prophase study.

[Bioaccumulation and toxicity test of diethylstilbestrol to zebrafish (Danio rerio) embryo].

The bioaccumulation regulation and the toxicity of diethylstilbestrol (DES) were studied on zebrafish (Danio rerio) embryos with the methods of HPLC and early life stage test, and the toxic mechanisms were also discussed with the techniques of quantitative structure-activity relationship (QSAR) and gene chips. After the zebrafish embryos exposed at 0.2 mg x L(-1) DES for 24, 48 and 72 h, the body levels of DES in zebrafish embryo were (25.78 +/- 4.95), (54.88 +/- 7.10) and (71.93 +/- 10.55) microg/g, respectively. Even exposure started at different time (0 hpf and 8 hpf), the most sensitive sub lethal endpoints were both 72 h delayed hatch, and the EC50 values for them were (0.14 +/- 0.09) and (0.33 +/- 0.14) mg/L, so that zebrafish embryo was more sensitive to the exposure of DES before the stage of gastrula (8 h after fertilization). QSAR studies found when compared with other phenol-endocrine disrupts, DES was much more toxic, and this related to its higher TE value (52.1217 eV). Meanwhile, when adult zebrafish exposed at 5 microg/L DES for 21 d, it was found by the gene chips that parts of gene expressions would be changed and the genetic toxicity of DES was approved. DES was concluded to be developmental and genetic toxic to zebrafish, and the biochemical reaction-associated process might be the key parameter in determining the toxicity level.

Herbal bioactivation: the good, the bad and the ugly.

It has been well established that the formation of reactive metabolites of drugs is associated with drug toxicity. Similarly, there are accumulating data suggesting the role of the formation of reactive metabolites/intermediates through bioactivation in herbal toxicity and carcinogenicity. It has been hypothesized that the resultant reactive metabolites following herbal bioactivation covalently bind to cellular proteins and DNA, leading to toxicity via multiple mechanisms such as direct cytotoxicity, oncogene activation, and hypersensitivity reactions. This is exemplified by aristolochic acids present in Aristolochia spp, undergoing reduction of the nitro group by hepatic cytochrome P450 (CYP1A1/2) or peroxidases in extrahepatic tissues to reactive cyclic nitrenium ion. The latter was capable of reacting with DNA and proteins, resulting in activation of H-ras oncogene, gene mutation and finally carcinogenesis. Other examples are pulegone present in essential oils from many mint species; and teucrin A, a diterpenoid found in germander (Teuchrium chamaedrys) used as an adjuvant to slimming diets. Extensive pulegone metabolism generated p-cresol that was a glutathione depletory, and the furan ring of the diterpenoids in germander was oxidized by CYP3A4 to reactive epoxide which reacts with proteins such as CYP3A and epoxide hydrolase. On the other hand, some herbal/dietary constituents were shown to form reactive intermediates capable of irreversibly inhibiting various CYPs. The resultant metabolites lead to CYP inactivation by chemical modification of the heme, the apoprotein, or both as a result of covalent binding of modified heme to the apoprotein. Some examples include bergamottin, a furanocoumarin of grapefruit juice; capsaicin from chili peppers; glabridin, an isoflavan from licorice root; isothiocyanates found in all cruciferous vegetables; oleuropein rich in olive oil; dially sulfone found in garlic; and resveratrol, a constituent of red wine. CYPs have been known to metabolize more than 95% therapeutic drugs and activate a number of procarcinogens as well. Therefore, mechanism-based inhibition of CYPs may provide an explanation for some reported herb-drug interactions and chemopreventive activity of herbs. Due to the wide use and easy availability of herbal medicines, there is increasing concern about herbal toxicity. The safety and quality of herbal medicine should be ensured through greater research, pharmacovigilance, greater regulatory control and better communication between patients and health professionals.