The Meridian Tropism and Classification of Red Yeast Rice Investigated by Monitoring Dermal Electrical Potential.

Red yeast rice is a traditional Chinese medicine and food that has been purported to color food, ferment, and lower cholesterol. In order to study the antioxidative capacity of red yeast rice and the effects on electrical potential difference (EPD) of 12 acupuncture meridians, the pH value, oxidation reduction potential (ORP), ABTS, FRAP, T-SOD, and particle size distribution of red yeast rice were analyzed. 20 volunteers were recruited and randomly divided into two groups, the red yeast rice group (10 g red yeast rice and 40 g water) and control CK group (50 g water). The left 12 acupuncture meridians' EPD was real-time monitored. Samples were taken at the 10th minutes. The whole procedure continued for 70 minutes. It is shown that the pH value of the red yeast rice was 4.22, the ORP was 359.63 mV, the ABTS was 0.48 mmol Trolox, the FRAP was 0.08 mmol FeSO4, the T-SOD was 4.71 U, and the average particle size was 108 nm (7.1%) and 398.1 nm (92.9%). The results of 12 acupuncture meridians' EPD showed that the red yeast rice can significantly affect the EPD of stomach, heart, small intestine, and liver meridians.

Effects of TAT-SOD at Acupoints on Essential Hypertension by Monitoring Meridians Electrical Potential.

OBJECTIVES: To investigate the effect on essential hypertension of the topical application of TAT-Cu, Zn-superoxide dismutase (TAT-SOD) at left acupoint Zusanli (ST 36), and to observe whether the change of electrical potential difference (EPD) can be related to the change of blood pressure. METHODS: Sixteen patients with essential hypertension and 16 healthy subjects were included in the study. EPD between the left acupoints of Yanglingquan (GB 34) and Qiuxu (GB 40) was firstly screened out for the EPD detection. An intracellular superoxide quenching enzyme, TAT-SOD, was topically applied to the acupoint ST 36 within an area of 1 cm2 once a day, and the influence on EPD was investigated. The dosage applied to TAT-SOD group (n=8) was 0.2 mL of 3000 U/mL TAT-SOD cream prepared by adding purified TAT-SOD to a vehicle cream, while placebo group (n=8) used the vehicle cream instead. The left acupoints of Yanglingquan (GB 34) and Qiuxu (GB 40) were selected for EPD measurement after comparing EPD readings between 5 acupoints on each of all 12 meridians. RESULTS: EPDs between the left acupoints of GB 34 and GB 40 for 16 patients of essential hypertension and 16 healthy subjects were 44.9±6.4 and 5.6±0.9 mV, respectively. Daily application of TAT-SOD for 15 days at ST 36 of essential hypertension patients significantly decreased systolic blood pressure (SBP) and diastolic blood pressure (DBP) of 179.6 and 81.5 mm Hg to 153.1 and 74.1 mm Hg, respectively. Responding to the change in blood pressure, EPD between the left acupoints of GB 34 and GB 40 also declined from 44.4 to 22.8 mV with the same trend. No change was observed with SBP, DBP and EPD between the left acupoints of GB 34 and GB 40 with the daily application of the placebo cream. CONCLUSION: Enzymatic scavenging of the intracellular superoxide at ST 36 proved to be effective in decreasing SBP and DBP. The results reconfirm the involvement of superoxide anions and its transportation along the meridians, and demonstrate that EPD between acupoints may be an indicator to reflect its functioning status. Moreover, preliminary results suggest a close correlation between EPD and blood pressure readings, implying a possibility of using EPD as a sensitive parameter for blood pressure and to monitor the effect of antihypertensive treatment.

A new formula to calculate activity of superoxide dismutase in indirect assays.

To calculate superoxide dismutase (SOD) activity rapidly and accurately by indirect SOD assays, a formula based on the ratio of the catalytic speed of SOD to the reaction speed of the indicator with superoxide anion was deduced. The accuracy of this formula was compared with the conventional formula based on inhibition in five indirect SOD assays. The new formula was validated in nearly the entire SOD activity range, whereas the conventional formula was validated only during inhibition of 40-60%. This formula might also be used for the assays of other enzymes.

Topical Application of TAT-Superoxide Dismutase in Acupoints LI 20 on Allergic Rhinitis.

Reactive oxygen species are products of cellular metabolism and assigned important roles in biomedical science as deleterious factors in pathologies. In fact, some studies have shown that the therapeutic benefits of taking antioxidants were limited and the potential for therapeutic intervention remains unclear. New evidences showed that ROS have some ability of intercellular transportation. For treating allergic rhinitis, as a novel intracellular superoxide quencher, TAT-SOD applied to acupoints LI 20 instead of directly to nasal cavity can be used to test that. TTA group apply TAT-SOD cream prepared by adding purified TAT-SOD to the vehicle cream to acupoints LI 20, while placebo group used the vehicle cream instead. TTN group applied the same TAT-SOD cream directly to nasal cavity three times daily. Symptom scores were recorded at baseline and days 8 and 15. For the overall efficacy rate, TTA group was 81.0%, while placebo group was 5.9% and TTN was 0%. Malondialdehyde levels decreased observably in TTA group, and superoxide dismutase, catalase, and glutathione peroxidase levels remained basically unaffected. Enzymatic scavenging of the intracellular superoxide at acupoints LI 20 proved to be effective in treating allergic rhinitis, while no improvement was observed with the placebo group and TTN group.

Monitoring the Effects of Acupoint Antioxidant Intervention by Measuring Electrical Potential Difference along the Meridian.

Previous studies suggest that superoxide anions are possibly traveling along acupuncture meridians. The electrical potential difference (EPD) between acupoints may be related to the movement. To test the above hypothesis, we conducted a study investigating the effects of acupoint antioxidant interventions on the meridian EPD. Firstly, ST39 (L) and ST44 (L) were screened out for the EPD detection along the stomach meridian, and ST36 (L) was selected for interventions including acumassage with the control cream, as well as the TAT-SOD cream for 30 minutes, or injection with reduced glutathione sodium. The EPD between ST39 and ST44 was recorded for 80 minutes and measured again 48 h later. While the EPD increased during the acumassage, the acumassage with TAT-SOD cream and the glutathione injection generated waves of EPD increased, indicating the migration or removal from the visceral organ of a greater quantity of superoxide. Remarkably lower EPD readings 48 h later with both antioxidant acupoint interventions than the mere acumassage imply a more complete superoxide flushing out due to the restored superoxide pathway at the acupoint after interventions. The results confirm superoxide transportation along the meridians and demonstrate a possibility of acupoint EPD measurement as a tool to monitor changes in the meridians and acupoints.

High-performance ion exchange chromatography of intact bacterial cells in the manner of molecules: 1. Establishment of methodology.

The feasibility of separating intact bacterial cells by high-performance ion exchange chromatography typical for macromolecular study was investigated. A common HPLC system coupled with a light scattering detector was employed; TOYOPEARL SuperQ-650C, a beaded anion exchanger, was used as the column media; bacterial cell samples were prepared by suspending colonies in the equilibrium buffer. Piperazine-hydrochloric acid buffer (0.02 M, pH 7.0) was chosen to be the equilibrium buffer after screening different buffer systems. The absorbed cells were eluted by a linear gradient of NaCl from 0 to 1.00 M in the equilibrium buffer within 30 min, at a flow rate of 1.0 mL/min. A distinctive chromatographic profile with high reproducibility and accuracy was obtained with all of the six kinds of bacteria tested. The eluted fractions for each sample were confirmed by microscopic analysis and physiological or biochemical identification to the bacterial cells applied. More importantly, the eluted cells retained full viability. It is apparent that living bacteria cells exhibited similar behaviors and properties with molecules in anion exchange chromatography, implying that chromatographic methods can become an effective approach in the studies of intact bacterial cells.

[Characterization of Ralstonia solanacearum in different growth phases by high performance ion exchange liquid chromatography analysis].

High performance ion exchange liquid chromatography and laser light scattering instrument were employed to characterize Ralstonia solanacearum in different growth phases. The pure culture of Ralstonia solanacearum was successfully separated into three characteristic fractions. Chromatographic behaviors of Ralstonia solanacearum in lag phase, logarithmic phase and stationary phase were carefully investigated, and their relationships to the cell concentration, pH of fermentation broth and extracellular polysaccharide (EPS I ) content of cell surface were analyzed. It was found that the majority of Ralstonia solanacearum cells obtained at 8h culture time could not be absorbed to the resin due to the strong motility of the cells which could apparently overcome the electrostatic interaction. Furthermore, when the mobility of the cells was destroyed by 4% formaldehyde treatment, the prolonged retention time was recorded due to the stronger adsorbility to the resin. On the other hand, the EPS I content of cell surface was determined to increase with the culture time, indicating an increasing interaction between the cells and the resin. EPS I content of three characteristic chromatographic fractions was determined, and it was found that the higher EPS I content led to the longer retention time of the fraction, which confirmed the above-mentioned observation. As a result, it is concluded that the formation of three chromatographic fractions of the pure culture of Ralstonia solanacearum is attributed to bacterial motility and the interaction of EPS I of cell surface with the anionic exchange resin, and the novel method of ion exchange chromatography of the intact bacterial cells can be a useful tool in bacteriological study.

[Mechanism study of anti-influenza effects of Radix Isatidis water extract by red blood cells capillary electrophoresis].

OBJECTIVE: To research the mechanism of the inhibition effects of BWE on cell attachment of influenza virus by capillary electrophoresis. METHOD: The morphologic difference of red cells after treating with BWE infected by influenza virus was detected with microscope, capillary electrophoresis and HA. RESULT: The pretreatment of the normal cells with BWE inhibited the attachment of influenza to the cells, while no meaningful inhibition was observed when influenza virus was pretreated before being inoculated to cells. CONCLUSION: The results indicate that the inhibition effects of BWE on cell attachment of influenza virus may be an important mechanism of anti-influenza activity of Radix Isatidis Extracts.

[The modulation of pilose antler extract (PAE) on the proliferation of rat osteogenic cells UMR-106].

OBJECTIVE: To investigate the modulation of pilose antler extract (PAE) on rat osteogenic cells UMR-106 in vitro. METHOD: Component P2 of PAE was isolated by Sephacryl S-200HR gel filtration chromatography. The proliferative effects of P2 and other components isolated by Sephacryl S-200HR on UMR-106 cells were investigated by MTT assay. RESULT: The P2 could significantly increase the proliferation rate of osteogenic cells. When the protein concentration of P2 was between 0.972 mg x L(-1) and 97.2 mg x L(-1), it could inhibit the proliferation of UMR-106 cells. But while the concentration was equal to or greater than 97.2 mg x L(-1), the P2 could increase the proliferation rate of cells, especially 477.92% at 9.72 g x L(-1), which was approximated to 499.62% of PAE. The molecular weight of the P2 was about 59 kDa determined by SDS-PAGE. On the other hand, inhibition was also observed in the sample of the P3, P4 and P5. CONCLUSION: Those regulative factors in PAE which have different molecular weight can affect the proliferation of UMR-106 cells two-wayly. And this adjustment also relies on the dose of those factors. This finding may help us to understand the possible mechanism of Chinese traditional medicine from animal materials.

[Significance and expression of vascular endothelial growth factor-C (VEGF-C) in esophageal squamous carcinoma and glioma].

BACKGROUND & OBJECTIVE: The research in recent years has demonstrated that vascular endothelial growth factor-C (VEGF-C) is expressed in certain malignant tumor cells, and up to now VEGF-C is the only growth factor specific for lymphangiogenesis. The relationship between the VEGF-C expression in malignant tumor tissues and lymphatic metastasis is still scarcely reported. This study was designed to compare the VEGF-C expression in human esophageal squamous carcinoma and glioma, and then to analyze the relationship between the VEGF-C expression and tumor lymphatic metastasis. METHODS: The expression of VEGF-C antigen was detected with immunohistochemical method in 72 human esophageal squamous carcinomas (29 cases with lymph node metastasis,43 cases without lymph node metastasis) and 23 human gliomas (diagnosed pathologically as astrocytoma in grades I to IV), followed by the further analysis on the relationship between VEGF-C expression and clinicopathological parameters. RESULTS: The expression of VEGF-C antigen was not found in any gliomas, while the positive VEGF-C expression rate achieved 38.88%(28/72) in the human squamous carcinoma, with 62.07%(18/29) in the cases with lymph node metastasis, 23.26%(10/43) in the cases without lymph node metastasis, 58.82% (20/34) in the T2 and T3 of invasion depth cases, and 21.05%(8/38)in the T1 of invasion depth cases. CONCLUSION: The expression of VEGF-C antigen was significantly associated with lymph node metastasis and invasion depth in esophageal squamous carcinoma. VEGF-C may act as a key factor in the facilitation of lymphatic metastasis in esophageal squamous carcinoma.

Capillary electrophoresis of erythrocytes.

Capillary electrophoresis (CE) of erythrocytes from different sources under various conditions is reported in this paper. It was found that erythrocyte samples from sheep, duck, and human showed characteristic and reproducible elution peaks, and that the retention times of A-, B-, AB-, and O-type erythrocytes from human blood were distinctively different; even subtle differences, among individuals with the same blood type could be detected by CE. A strictly linear correlation was obtained between the peak area and the amount of human erythrocyte over a range of 4.8 x 10(2)-1.9 x 10(4) cells (r=0.999), indicating that CE could be used for rapid and accurate quantification of erythrocytes. Using this CE protocol, the decrease of the surface electrical charge of erythrocyte during storage was confirmed. Therefore, this work demonstrated that CE could be a useful alternative for characterizing and quantifying erythrocytes or other cells.

Expression, purification, and characterization of an unstable lysozyme mutant in Pichia pastoris.

To investigate the expression and purification of an unstable heterologous protein in Pichia pastoris, the cDNA of H5-lysozyme, a hen egg lysozyme mutant with a hydrophobic pentapeptide (Phe-Phe-Val-Ala-Pro) fused to the carboxyl terminus, was integrated into the genome of P. pastoris. It was found that medium composition, induction time, and fermenter type were important factors for the expression of H5-lysozyme. Substantially active H5-lysozyme was secreted by induction with methanol when the prepro-sequence of alpha-factor was used as secretion signal sequence. The amount secreted was 422-fold greater than that observed with Saccharomyces cerevisiae. Recombinant H5-lysozyme was recovered and purified by cation-exchange chromatography directly from fermentation broth. The mutant lysozyme showed bactericidal activity against Gram-positive as well as Gram-negative bacteria.