Experimental study of curcumol inhibiting vascular endothelial growth factor-induced neovascularization / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To study the role and mechanism of curcumol in neovascularization induced by vascular endothelial growth factor(VEGF).METHODS: Human umbilical vein endothelial cells were cultured in vitro and treated with 50ng/mL VEGF and curcumol at different concentrations. Cell proliferation was detected by CCK-8 and EdU assay, the migration ability of cells was analyzed by Transwell assay, the angiogenesis ability of endothelial cells was analyzed by tube formation assay, and the change of Akt/mTORC1 signal pathway was detected by Western blot.RESULTS: CCK-8 results showed that the OD450 value of cells in 400 and 800 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(all P<0.01). EdU results showed that the rate of cell proliferation in 400 μmol/L curcumol+VEGF group was significantly lower than that in VEGF group(P<0.001). Transwell assay and the formation assay results showed that the number of migratory cells in 400 μmol/L curcumol+VEGF group was decreased, and the number and length of tube branches were also reduced compared with VEGF group(all P<0.001). Western blot results showed that curcumol significantly inhibited the expression of p-Akt and p-S6, which were downstream targets of Akt/mTORC1 pathway in cells.CONCLUSION: Curcumol can inhibit VEGF-induced cell proliferation, migration and tube formation of vein endothelial cells, and has a strong inhibitory effect on angiogenesis, which can be further studied in the treatment of ocular fundus neovascularization.

Effect of Isopimpinellin on Expression of Inhibitory Neurotransmitter and Receptor Genes in Primary Hippocampal Neurons / 中国实验方剂学杂志

Objective:To observe the effect of isopimpinellin on primary hippocampal neuron cells γ-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT) and receptor genes expressions, in order to explore its hypnotic mechanism. Method:The primary hippocampal neurons of neonatal Sprague-Dawley rats were cultured in vitro. And subsequent experiments were conducted in the optimal state of cell growth, and the purity was identified by immunohistochemistry of neuron-specific enolase. Hippocampal neurons were randomly divided into five groups, namely blank control group, diazepam group (25 mg·L-1), and low-dose (5 mg·L-1), moderate-dose (10 mg·L-1) and high-dose (20 mg·L-1) isopimpinellin groups. Early apoptosis of hippocampus neuron cells were detected using flow cytometry technique after 24 h administration, and the changes in the levels of GABA and 5-HT were detected using enzyme-linked immunosorbent. The changes in mRNA expressions of receptor genes relating to gamma-aminobutyric acid type A receptor(GABAA) genes GABRA1,GABRA5,GABBR1, gamma-aminobutyric acid type B receptor genes (GABAB) GABRB2, 5-hydroxytryptamine 1A receptor (5-HT1A)5-HT1A(A),5-HT1A(B),5-HT1A(C) were detected by real-time quantitative PCR(Real-time PCR). Result:On the 7th day, the hippocampal neurons grew in a good condition, and the purity was above 90%. Apoptosis rates of hippocampal neurons in the low-dose and moderate-dose groups were significantly lower than that in the blank control group (PP1,GABRA5,5-HT1A(A),5-HT1A(C) in the moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (PP1,5-HT1A(B) in the low-dose, moderate-dose and high-dose isopimpinellin groups were significantly higher than those in the blank control group (PPConclusion:The hypnotic mechanism of isopimpinellin may be related to the inhibition of hippocampal neuron apoptosis, the increase of the content of inhibitory neurotransmitter GABA, and the up-regulation of GABA and 5-HT-related receptor genes.

The potential protein kinase A (Pka) phosphorylation site is required for the function of FgSge1 in Fusarium graminearum.

The new transcription factor Sge1 has garnered much attention in filamentous fungi recently, which highlights its role in pathogenicity, conidiation, and the production of secondary metabolites. In this study, we demonstrated that FgSge1 is localized in the nucleus in Fusarium graminearum using fluorescent protein GFP. Mutants containing a T67A mutation within the potential protein kinase A (Pka) phosphorylation site of FgSge1 exhibited a significant decrease in conidiation and dramatically impaired virulence on both wheat head and non-host tomato. These results indicated that the Pka phosphorylation site is required for the function of FgSge1 in F. graminearum. In addition, we characterized the FgSGE1 deletion mutants and found that the mutants showed increased sensitivity to osmotic stress mediated by NaCl or KCl, and to cell wall damaging agent congo red (CR). Real-time PCR assays revealed increased transcription levels of FgSGE1 with the treatment of NaCl or CR, and decreased FgSGE1 transcription in the FgOS-2 deletion mutant ΔFgOs-2. Based on the transcription levels, it can be concluded that FgSge1 is a downstream target of the mitogen-activated protein kinase FgOs-2.

[Application of thermal dissipation probe in the study of Bambusa chungii sap flow].

Based on the validation of Granier's empirical formula for calculating tree stem sap flux density, a comparative study was conducted on the measurement of Bambusa chungi sap flow by using different lengths of thermal dissipation probe (TDP), aimed to approach the applicability of TDP in measuring the sap flow of B. chungii. The difference in the daily change of the sap flow between B. chungii and nearby growing Schima superb was also analyzed. Because of the thinner bamboo wall and the heterogeneous anatomy, the sap flux density of B. chungii measured by 10 mm long probe could be underestimated, but that measured by 8 and 5 mm long probes could be relatively accurate. The comparison of the sap flow between B. chungii and nearby growing S. superba revealed that both the mean sap flux density and its daily change pattern' s skewness of B. chungii were higher than those of S. superba, but the nighttime sap flow of B. chungii was less than that of S. superba, indicating that the water recharge of B. chungii during nighttime was less active than that of S. superba. It was suggested that using TDP to investigate the sap flow of bamboo would be feasible, but careful calibration would be required before the TDP was put into application on different bamboo species.

[Effects of tree diameter at breast height and soil moisture on transpiration of Schima superba based on sap flow pattern and normalization].

The eigenvalues of continuous sap flow pattern, i. e. , skewness and kurtosis, were used to investigate the water usage of Schima superba with different diameter at breast height (DBH), and the method of normalization was firstly applied to eliminate the effects of strong affecting factor (photosynthetic active radiation, PAR) to explore the possible relationship between weak affecting factor (soil moisture) and sap flow. Generally, the trees with larger DBH had smaller skewness of sap flux density and later-appeared but larger peak values, suggesting that much more water was transpired, and the larger trees showed smaller skewness and later-appeared larger peak values in wet season than in dry season, suggesting that more water was transpired in wet season. On the other hand, smaller trees had lesser differences in the skewness between dry and wet seasons, suggesting that there was no significant difference in the transpiration between the two seasons. The relationship between individual tree's transpiration and soil moisture was significant and positive after the two parameters being normalized with PAR peak values. When the soil moisture content was higher, the transpiration of the trees with larger DBH was steadily increasing with soil moisture, while that of the trees with moderate or smaller DBH had opposite trend, presumably due to their transpiration and water absorption were approached to the limit.