[Analyze moisture transformation and transport rules during processing of Paeoniae Radix Alba by using low-field NMR].

Moisture status and content during the processing of Paeoniae Radix Alba were studied by nuclear magnetic resonance (NMR) and nuclear magnetic resonance imaging (MRI) to investigate the changes of transverse relaxation time (T 2) and MRI images during boiling and drying processes of Paeoniae Radix Alba. The results showed that water in Paeoniae Radix Alba fresh products was major of free water, and in the boiling process, the content of free water increased whereas the content of bound water declined. At the end of boiling, content of free water reached over 90%. During the drying process, T 2 moved to the left, and moisture mobility was reduced. The MRI image directly showed that moisture transfer was outside-in process for both increase and decrease. At the end of drying, remaining moisture was mainly present in inner layer of Paeoniae Radix Alba. Quality and appearance were affected by the change of moisture during processing process of medicinal herbs. NMR and MRI could provide direct reference evidence for its moisture changes, and the results of this study could provide direct references and technical support for optimization of processing process of root medicinal materials and evaluation of Chinese herbal pieces.

[Chemical constituents from flowers of Paeonia lactiflora].

OBJECTIVE: To study the chemical constituents of Paeonia lactiflora flowers. METHODS: [corrected] The chemical constituents were isolated and purified by various chromatography methods,and the structures were identified by physicochemical and modem spectroscopic. RESULTS: 11 compounds were identified as gallic acid(1),methyl gallate(2),ethyl gallate(3),1,2,3,6-tetragalloyl-beta-D-glucopyranoside(4), 1,2,3,4,6-pentagalloyl-beta-D-glucopyranoside(5), quercetin-3-O-glucoside-6"-gallate(6), kaempferol-3-O-glucoside-6"-gallate(7), 1-O-galloyl-beta-D-glucose (8), kaempferol-3, 7-di-O-beta-D-glucoside(9), paeoniflorin(10) and albiflorin(11). CONCLUSIONS: Compounds 1-8, 10 and 11 are obtained from the flowers of Paeonia lactiflora for the first time,compounds 6 and 7 are obtained from Paeonia genus for the first time.

MicroRNA-29b inhibits peritoneal fibrosis in a mouse model of peritoneal dialysis.

TGF-ß/Smad3 signaling plays a pivotal role in the pathogenesis of peritoneal fibrosis associated with peritoneal dialysis (PD). MicroRNA-29 (miR-29) is known as a potent downstream inhibitor of TGF-ß/Smad3 in renal fibrosis. In this study, we examined the therapeutic potential for miR-29b on PD-related peritoneal fibrosis in a mouse model of PD induced by daily infusion of 4.25% dextrose-containing PD fluid (PDF). MiR-29b-expressing plasmid was delivered into the peritoneum via an ultrasound-microbubble-mediated system before and at day 14 after PDF. We found that mice on PD developed peritoneal fibrosis with impaired peritoneal function, which was associated with a loss of miR-29b. In contrast, overexpression of miR-29b before the PDF infusion showed a protective effect on peritoneal fibrosis including EMT and prevented peritoneal dysfunction. Moreover, delayed miR-29b treatment until peritoneal fibrosis was established at day 14 also halted the progression of peritoneal fibrosis at day 28. Further studies identified that blockade of the Sp1-TGF-ß/Smad3 pathway may be a mechanism by which miR-29b inhibited peritoneal fibrosis. In conclusion, treatment with miR-29b may represent a novel and effective therapy for PD-associated peritoneal fibrosis.

Opposing roles for Smad2 and Smad3 in peritoneal fibrosis in vivo and in vitro.

Peritoneal fibrosis is a major cause of ultrafiltration failure in patients receiving continuous ambulatory peritoneal dialysis. Transforming growth factor (TGF)-ß1 is an important mediator in this process; however, its signaling mechanisms had not been explored. Thus, we examined TGF-ß1/Smad signaling in human peritoneal biopsy specimens associated with continuous ambulatory peritoneal dialysis. We found that TGF-ß/Smad2/3 signaling was highly activated in patients with increased collagen deposition and thickening of the peritoneal membrane who were receiving continuous ambulatory peritoneal dialysis. Long-term exposure of wild-type mice to 4.25% peritoneal dialysis solution for 30 days induced significant peritoneal fibrosis with impaired peritoneal equilibrium, which was prevented in Smad3 knockout mice. In contrast, conditional Smad2 gene deletion in the peritoneum exacerbated peritoneal fibrosis and dysfunction. The contrasting roles of Smad2 and Smad3 in peritoneal fibrosis were also examined in vitro. Cultured mesothelial cells from Smad3 knockout mice were resistant to TGF-ß1-induced collagen I production and the transition toward a myofibroblast phenotype as seen in wild-type cells, whereas Smad2 deficiency in mesothelial cells failed to modulate the profibrotic response to TGF-ß1. In conclusion, this study found activation of TGF-ß/Smad signaling in peritoneal fibrosis in patients receiving continuous ambulatory peritoneal dialysis and identifies opposing roles for Smad2 and Smad3 in peritoneal dialysis-associated peritoneal fibrosis. These findings provide a mechanistic basis for future therapies targeting TGF-ß/Smad signaling in peritoneal fibrosis.

Monoterpenes from Paeonia albiflora and their inhibitory activity on nitric oxide production by lipopolysaccharide-activated microglia.

Eleven new monoterpenes, paeonidangenin (1), paeonidanin A (2), paeonidanin B (3), paeonidanin C (4), paeonidanin D (5), paeonidanin E (6), paeoniflorone (7), 4-O-methylbenzoylpaeoniflorin (8), 4-O-methylgalloylpaeoniflorin (9), 4-O-methyldebenzoylpaeoniflorin (10), and 4-O-methylalbiflorin (11), were isolated from the 60% ethanol extract of the roots of Paeonia albiflora. Their structures were determined primarily on the basis of 1D and 2D NMR techniques and MS studies. Paeonidanins D (5) and E (6) are unprecedented examples of "cage-like" monoterpene dimers. The inhibitory effects of the isolated compounds on nitric oxide production by lipopolysaccharide (LPS)-activated N9 microglia were evaluated.

Four new compounds from Paeonia albiflora.

Studies on the chemical constituents of the roots of Paeonia albiflora Pall. led to the isolation of four new compounds named (3R,4S)-3-methyl-3,4-dihydro-5,6,7-trihydroxy-4-(3'-methoxy-4'-hydroxyphenyl)-1H-[2]-benzopyran-1-one (1), 5-hydroxy-6-methyl-1H-indole-3-carbaldehyde (2), trans-5-hydroxy-2-methoxy-6-methyl-2,3-dihydrobenzofuran-3-yl methyl benzoate (3) and cis-5-hydroxy-2-methoxy-6-methyl-2,3-dihydrobenzofuran-3-yl methyl benzoate (4), and two known ones, (7S,8S)-3-methoxy-3',7-epoxy-8,4'-oxyneligna-4,9,9'-triol (5) and (7S,8R)-dihydrodehydrodiconifery alcohol (6). Their structures were determined mainly by spectroscopic techniques including 2D-NMR (HSQC, HMBC, NOESY), MS, and CD experiments.

[Urine protein from patients with lupus nephritis stimulates transdifferentiation and high expression of collagen in renal tubular cells].

OBJECTIVE: To investigate the effects of urine protein on the renal tubular-interstitial fibrosis in the patients with lupus nephritis (LN). METHODS: Protein was isolated and purified from the urine of six patients with primary LN, 1 male and 5 females, aged 27.4, and incubated with renal tubular cells of the line HK-2 for 0, 1, 2, 12, 24, or 48 h respectively. The mRNA expressions of transforming growth factor beta1(TGF-beta1), collagen I (COL I), and alpha-smooth muscle actin (alpha-SMA) in the HK-2 cells were detected by RT-PCR, and the. protein expressions of TGF-beta1, COL I, and alpha-SMA were detected with Western blotting and indirect immunofluorescence. RESULTS: The urine protein from the LN patients dose-and time-dependently increased the mRNA and protein expressions of TGF-beta1, COL I, and alpha-SMA in the HK-2 cells. The TGF-beta1 mRNA level 48 h after incubation was 0.39 +/- 0.03, significantly higher than that at the beginning of incubation (0.27 +/- 0.02, P < 0.01), and the TGF-beta1 protein level 48 h after incubation was 0.37 +/- 0.03, 1.7 times that at the beginning of incubation (0.27 +/- 0.04, P < 0.01). The COL I mRNA level 48 h after incubation was 0.38 +/- 0.02, significantly higher than the baseline level (0.22 +/- 0.03, P < 0.01); and the COL I protein level 48 h after incubation was 0.44 +/- 0.03, significantly higher than the baseline level (0.19 +/- 0.02, P < 0.01). The alpha-SMA mRNA level 48 h after incubation was 0.66 +/- 0.04, significantly higher than the baseline level (0.44 +/- 0.03, P < 0.01), and the alpha-SMA protein level 48 h after incubation was 0.43 +/- 0.02, significantly higher than the baseline level (0.24 +/- 0.03, P < 0.01). CONCLUSION: Urine protein may play an important role in the renal tubular-interstitial fibrosis by inducing the production of extracellular matrix and phenotype change in HK-2 cells.