Bipolar Membrane Electrodialysis for Direct Conversion of L-Ornithine Monohydrochloride to L-Ornithine.

In this study, bipolar membrane electrodialysis was proposed to directly convert L-ornithine monohydrochloride to L-ornithine. The stack configuration was optimized in the BP-A (BP, bipolar membrane; A, anion exchange membrane) configuration with the Cl- ion migration through the anion exchange membrane rather than the BP-A-C (C, cation exchange membrane) and the BP-C configurations with the L-ornithine+ ion migration through the cation exchange membrane. Both the conversion ratio and current efficiency follow BP-A > BP-A-C > BP-C, and the energy consumption follows BP-A < BP-A-C < BP-C. Additionally, the voltage drop across the membrane stack (two repeating units) and the feed concentration were optimized as 7.5 V and 0.50 mol/L, respectively, due to the low value of the sum of H+ ions leakage (from the acid compartment to the base compartment) and OH- ions migration (from the base compartment to the acid compartment) through the anion exchange membrane. As a result, high conversion ratio (96.1%), high current efficiency (95.5%) and low energy consumption (0.31 kWh/kg L-ornithine) can be achieved. Therefore, bipolar membrane electrodialysis is an efficient, low energy consumption and environmentally friendly method to directly convert L-ornithine monohydrochloride to L-ornithine.

[Improvement of survival quality of the patients with hemodialysis treated with moxibustion for regulating spleen and stomach functions: multi-central randomized controlled study].

OBJECTIVE: To discuss the impacts of moxibustion for regulating spleen and stomach function on the survival quality of the patients of end stage renal disease (ESRD) with maintenance hemodialysis (MHD). METHODS: One hundred and nine cases of uremia with MHD from 3 hemodialysis centers were randomized into an observation group (58 cases) and a control group (51 cases). The regular hemodialysis and conventional medication were used in the two groups. In the observation group, on the basis of the common treatment, moxibustion was applied to Zusanli (ST 36) and Sanyinjiao (SP 6), 2-3 times a day, the treatment of 4 weeks made one session. Totally, 3 sessions were required and the follow-up lasted for 3 months. KDQOL-SF (kidney disease quality of life short form,KDQOL-SFTM 1. 3) was adopted for the questionnaire investigation on survival quality before treatment, after treatment and at the end of follow-up separately in the two groups. RESULTS: After treatment, the survival quality scores in terms of physical functioning (83.62+/-13.27 vs 79.32+/- 22. 17), general health (58. 88+/- 20.24 vs 48.82+/-20.89) and vitality (77.07+/-15.56 vs 70. 59+/-22.61) in the observation group were higher than those in the control group (all P<0. 05). In comparison before and after treatment in the same group, the survival quality scores in terms of physical functioning, general health, vitality and symptoms/problems were all improved in the observation group (all P<0. 05). At the end of follow-up, the survival quality scores in terms of physical functioning, general health, mental health, social functioning, vitality, effects of kidney disease and cognitive function were higher in the observation group as compared with those in the control group (all P<0. 05). In comparison of the results at the end of follow-up with those before treatment, the survival quality scores in terms of vitality, symptoms/problems and cognitive function in the observation group were improved (all P< 0. 05). The differences were not significant in all of the 19 fields of survival quality evaluation before and after treatment, and after follow-up in the control group (all P>0. 05). CONCLUSION: Moxibustion for regulating spleen and stomach function improves the survival quality of the patients with hemodialysis in terms of physical functioning, general health and vitality, which benefits the psychological condition of the patients, resulting in the improvements of the survival quality in the fields of mental health, social functioning, effects of kidney disease and cognitive function.

Induction of DNA damage and p21-dependent senescence by Riccardin D is a novel mechanism contributing to its growth suppression in prostate cancer cells in vitro and in vivo.

PURPOSE: Our previous studies had shown that Riccardin D (RD) exhibited cytotoxic effects by induction of apoptosis and inhibition of angiogenesis and topoisomerase II. Here, we reported that apoptosis is not the sole mechanism by which RD inhibits tumor cell growth because low concentrations of RD caused cellular senescence in prostate cancer (PCa) cells. METHODS: Low concentrations of RD were used to treat PCa cells in vitro and in vivo, and senescence-associated ß-galactosidase activity, DNA damage response markers, and/or colony-forming ability, cell cycle were analyzed, respectively. We then used siRNA knockdown to identify key factor in RD-triggered cellular senescence. RESULTS: RD treatment caused growth arrest at G0/G1 phase with features of cellular senescence phenotype such as enlarged and flattened morphology, increased senescence-associated-beta-galactosidase staining cells, and decreased cell proliferation in PCa cells. Induction of cellular senescence by RD occurred through activation of DNA damage response including increases in the phosphor-H2AX, inactivation of Chk1/2, and suppression of repair-related Ku70/86 and phosphor-BRCA1 in PCa cells in vitro and in vivo. Analysis of expression levels of p53, p21(CIP1), p16(INK4a), p27(KIP1), pRb and E2F1 and genetic knockdown of p21(CIP1) demonstrated an important role of p21(CIP1) in RD-triggered cellular senescence. CONCLUSIONS: Involvement of the DNA damage response and p21(CIP1) defines a novel mechanism of RD action and indicates that RD could be further developed as a promising anticancer agent for cancer therapy.

[Randomized controlled trail on moxibustion for maintenance hemodialysis patients in deficiency syndrome].

OBJECTIVE: To explore the effective therapy for maintenance hemodialysis patients in dificiency syndrome in end-stage renal disease. METHODS: Ninety-seven cases were divided into an observation group (51 cases) and a control group (46 cases) randomly, and routine western medicine was used in both of them. On this base, moxibustion was used in Zusanli (ST 36) and Sanyinjiao (SP 6) in paper-tube-moxibustion equipment in the observation group. Evaluate the therapeutic effect on symptoms by comparing the symptom scores in two groups before and after treatment. RESULTS: All the symptom scores in the observation group were improved after treatment, and the differences were significant (all P < 0.05). Among all symptoms, the most improved ones included lassitude and fatigue, short breath and aversion to talk, poor appetite, soreness and softness of waist and knees, aversion to cold, cold extremities and so on. In the aspect of therapeutic effect on symptoms, the total effective rate in observation group (64.7%, 33/51) was higher than that in control group (23.9%, 11/46), and the difference was significant (P < 0.05). CONCLUSION: Moxibustion can improve the clinical symptoms of maintenance hemodialysis patients in end-stage renal disease, and can generate some therapeutic effect to the dificiency syndrome of this disease.

Enhancement of matrix production and cell proliferation in human annulus cells under bioreactor culture.

Tissue engineering is a promising approach for treatment of disc degeneration. Herein, we evaluated effects of rotating bioreactor culture on the extracellular matrix production and proliferation of human annulus fibrosus (AF) cells. AF cells were embedded into alginate beads, and then cultured up to 3 weeks in a rotating wall vessel bioreactor or a static vessel. By real-time reverse transcription-polymerase chain reaction, expression of aggrecan, collagen type I and type II, and collagen prolyl 4-hydroxylase II was remarkably elevated, whereas expression of matrix metalloproteinase 3 and a disintegrin and metalloproteinase with thrombospondin motifs 5 was significantly decreased under bioreactor. Biochemical analysis revealed that the levels of the whole cell-associated proteoglycan and collagen were approximately five- and twofolds in rotating bioreactor, respectively, compared to those in static culture. Moreover, AF cell proliferation was augmented in rotating bioreactor. DNA contents were threefolds higher in rotating bioreactor than that in static culture. Expression of the proliferating cell nuclear antigen was robustly enhanced in rotating bioreactor as early as 1 week. Our findings suggested that rotating bioreactor culture would be an effective technique for expansion of human annulus cells for tissue engineering driven treatment of disc degeneration.

Ginsenoside Rg3 promotes beta-amyloid peptide degradation by enhancing gene expression of neprilysin.

OBJECTIVES: It has been hypothesized that the accumulation of beta-amyloid peptide (Abeta) in the brain is a triggering event leading to the pathological cascade of Alzheimer's disease. The steady-state levels of Abeta are determined by the metabolic balance between anabolic and catabolic activity and the dysregulation of this activity leads to Alzheimer's disease. Recent evidence has shown that neprilysin (NEP) is the rate-limiting enzyme in the Abeta degradation in the brain. Ginseng, the root of Panax ginseng C.A. Meyer, is widely used as a tonic for the prevention and treatment of age-related disorders in China. We aimed to investigate the basis of this use. METHODS: In this study, we investigated the effect of ginsenoside Rg3, one of the major active components of ginseng, on the metabolism of Abeta40 and Abeta42 in SK-N-SH cells transfected with Swedish mutant beta-amyloid precursor protein (SweAPP). RESULTS: The ELISA result showed that Rg3 significantly reduced the levels of Abeta40 and Abeta42, 19.65 +/- 6.05%, 23.61 +/- 6.74%, respectively (P < 0.01). The Western blot analysis showed that Rg3 reduced the levels of Abeta40 and Abeta42 through enhancing NEP gene expression, and real-time PCR assay showed that 50 microM Rg3 could significantly enhance NEP gene expression (2.9 fold at 48 h). CONCLUSIONS: Our findings suggest that the Rg3 compound of ginseng may be useful for treating patients suffering with Alzheimer's disease.

Enhancement of neutral endopeptidase activity in SK-N-SH cells by ginsenoside Rb1.

OBJECTIVES: To investigate the effect of ginsenosides Rb1 and Rg1 on Neprilysin (NEP) activity in SK-N-SH cells, and probe the underlying mechanism. METHODS: The effects of ginsenosides Rb1 and Rg1 on NEP activity were analyzed by NEP peptidase assay. Western blot was used to determine NEP gene expression at translational level, and RT-PCR was also performed to detect NEP gene expression at transcriptional level. RESULTS: NEP peptidase assay indicated that ginsenoside Rb1 can improve the activity of NEP, and RT-PCR and western blot results showed that the enhancement of NEP activity by ginsenoside Rb1 was due to enhancing NEP gene expression, while Rg1 did not have this effect. CONCLUSION: Our studies showed that ginsenoside Rb1 can enhance NEP activity by upregulating NEP gene expression. Our findings might offer a pharmacological explanation for the use of ginseng in traditional medicine.

Membrane-bound L- and D-lactate dehydrogenase activities of a newly isolated Pseudomonas stutzeri strain.

Pseudomonas stutzeri SDM was newly isolated from soil, and two stereospecific NAD-independent lactate dehydrogenase (iLDH) activities were detected in membrane of the cells cultured in a medium containing DL-lactate as the sole carbon source. Neither enzyme activities was constitutive, but both of them might be induced by either enantiomer of lactate. P. stutzeri SDM preferred to utilize lactate to growth, when both L-lactate and glucose were available, and the consumption of glucose was observed only after lactate had been exhausted. The Michaelis-Menten constant for L-lactate was higher than that for D-lactate. The L-iLDH activity was more stable at 55 degrees C, while the D-iLDH activity was lost. Both enzymes exhibited different solubilization with different detergents and different oxidation rates with different electron acceptors. Combining activity staining and previous proteomic analysis, the results suggest that there are two separate enzymes in P. stutzeri SDM, which play an important role in converting lactate to pyruvate.

Pseudomonas stutzeri as a novel biocatalyst for pyruvate production from DL-lactate.

Pseudomonas stutzeri SDM oxidized DL-lactic acid (25.5 g l(-1)) into pyruvic acid (22.6 g l(-1)) over 24 h. Both NAD(+)-independent D-lactate dehydrogenase and NAD(+)-independent L-lactate dehydrogenase were found for the first time in the bioconversion of lactate to pyruvate based on the enzyme activity assay and proteomic analysis.