Role of L-Arginine on the Expression of Coagulation Factor VIII Gene.

Objective: To explore the key sites in which L-arginine affects the expression of human coagulation factor VIII gene, and to create new drug targets for the treatment of hemophilia. Methods: A total of 5 human FVIII genes (A1, A2, A3, C1 and C2) with B domain deletion were transfected into human umbilical vein endothelial cells (HUVECs) as promoters. Run-on assay and ELISA analysis were performed to observe the driving effect of each domain gene on chloramphenicol acetyl transferase (CAT) gene transcription and expression, and the effect of L-arginine on each promoter. Results: In co-culture with L-arginine, transcriptional expression of the CAT gene was not detected in the PCAT3-Basic group (negative control without promoters), PA3-CAT3-Enhancer group or PC1-CAT3-Enhancer group. The transcriptional expression of CAT gene in the PCAT3-Control group (positive control with promoters) and PA1-CAT3-Enhancer group was unchanged compared with the non-L-arginine intervention, while the transcriptional expression of CAT gene in the PA2-CAT3-Enhancer group was significantly enhanced. Conclusions: A1 and A2 domain genes had promoter function and could initiate the transcription and expression of CAT gene, but A3, C1 and C2 domain genes could not. Moreover, L-arginine can significantly enhance transcription and expression of human coagulation factor VIII via A2 domain.

[Effect of Aerobic/Phosphorus Granules on Start-up of Partial Nitrification Granular Sludge].

The different effects of additional aerobic granules (AGs) and phosphorus removal granules (PRGs) on the start-up and stable operation of partial nitrification granular sludge (PNGS) were compared at room temperature(22-28℃). The results showed that in the first stage (days 0-22), partial nitrification was accomplished on day 19 for the three reactors (R1, R2, and R3). In the second stage (days 23-56), 20% AGs and 20% PRGs were added to R2 and R3 to induce PNGS. The start-up of the granules of the three reactors was successfully achieved. The mean particle sizes of R1, R2, and R3 reached 412 µm at day 76, 468 µm at day 42, and 400 µm at day 56. In the third stage (days 57-108), because the influent ammonia load increased from 0.4 kg·(m3·d)-1 to 0.5 kg·(m3·d)-1 and the COD load increased from 0.2 kg·(m3·d)-1 to 0.5 kg·(m3·d)-1, the mean particle sizes of R1 and R2 increased significantly. The average particle sizes of R1 and R2 reached 689 µm and 893 µm by the end of the operation (day 108), but sludge expansion occurred in R3. The inoculation of either AGs or PRGs could quickly achieve granulation, but the PNGS inoculated with the AGs could adapt to higher C/N and be more tolerable to shock loads and long-term stable operation.

Stachydrine, a major constituent of the Chinese herb leonurus heterophyllus sweet, ameliorates human umbilical vein endothelial cells injury induced by anoxia-reoxygenation.

Stachydrine is a major constituent of Chinese herb leonurus heterophyllus sweet, which is used in clinics to promote blood circulation and dispel blood stasis. Our study aimed to investigate the role of stachydrine in human umbilical vein endothelial cells (HUVECs) injury induced by anoxia-reoxygenation. Cultured HUVECs were divided randomly into control group, anoxia-reoxygenation (A/R) group and 4 A/R+stachydrine groups. HUVECs in the control group were exposed to normoxia for 5 hours, while in all A/R groups, HUVECs underwent 3 hours anoxia followed by 2 hours reoxygenation, and HUVECs in the 4 A/R+stachydrine groups were treated with 10(-8) M, 10(-7) M, 10(-6) M and 10(-5) M (final concentration) of stachydrine respectively. After anoxia-reoxygenation, tissue factor (TF) was over-expressed, cell viability and the concentrations of SOD, GSH-PX and NO were declined, while LDH, MDA and ET-1 were over-produced (p < 0.05 to 0.001 vs. the control group). However, in stachydrine treated groups, TF expression was inhibited at both mRNA and protein levels, while the declined cell viability and SOD, GSH-PX, NO as well as the enhanced LDH, MDA and ET-1 levels occurred during anoxia-reoxygenation were ameliorated and reversed effectively (p < 0.05 to 0.01 versus A/R group). Consequently, our findings indicate that TF plays an important role in the development of anoxia-reoxygenation injury of HUVECs, stachydrine ameliorates HUVECs injury induced by anoxia-reoxygenation and its putative mechanisms are related to inhibition of TF expression.