Growth under elevated air temperature alters secondary metabolites in Robinia pseudoacacia L. seedlings in Cd- and Pb-contaminated soils.

Plant secondary metabolites play a pivotal role in growth regulation, antioxidant activity, pigment development, and other processes. As the global climate changes, increasing atmospheric temperatures and contamination of soil by heavy metals co-occur in natural ecosystems, which alters the pH of rhizosphere soil and influences the bioavailability and mobility of metals. Elevated temperatures in combination with heavy metals are expected to affect plant secondary metabolites, but this issue has not been extensively examined. Here, we investigated secondary metabolites in Robiniapseudoacacia seedlings exposed to elevated temperatures using a passive warming device in combination with Cd- and Pb-contaminated soils. Heavy metals significantly stimulated the accumulation of saponins, phenolic compounds, and flavonoids in leaves and stems; alkaloid compounds increased in leaves and decreased in stems, and condensed tannins fluctuated. Elevated temperatures, alone and in combination with Cd and Pb, caused increases in secondary metabolites in the plant tissues. Phenolic compounds showed the greatest changes among the secondary metabolites and significant interactive effects of temperature and metals were observed. These results suggest that slightly elevated temperature could enhance protective and defense mechanisms of Robinia pseudoacacia seedlings exposed to heavy metals by stimulating the production of secondary metabolites.

Salmonella induce autophagy in melanoma by the downregulation of AKT/mTOR pathway.

Salmonella have been demonstrated to inhibit tumor growth. However, the mechanism of Salmonella-induced tumor cell death is less defined. Autophagy is a cellular process that mediates the degradation of long-lived proteins and unwanted organelles in the cytosol. Tumor cells frequently display lower levels of basal autophagic activity than their normal counterparts and fail to increase autophagic activity in response to stresses. Autophagy is involved in the cell defense elimination of bacteria. The signaling pathways leading to activation of Salmonella-induced autophagy in tumor cells remain to be elucidated. We used autophagy inhibitor (3-Methyladenine) and apoptosis inhibitor (Z-VAD-FMK) to demonstrate that Salmonella may induce cell death via apoptosis and autophagic pathway. Meanwhile, we suggested that Salmonella induce autophagy in a dose- and time-dependent manner. The autophagic markers were increased after tumor cell infected with Salmonella. In addition, the protein express levels of phosph-protein kinase B (P-AKT), phosph-mammalian targets of rapamycin (P-mTOR), phosph-p70 ribosomal s6 kinase (P-p70s6K) in tumor cells were decreased by western analysis after Salmonella infection. In conclusion, our results point out that Salmonella induce the autophagic signaling pathway via downregulation of AKT/mTOR pathway. Herein, our findings that Salmonella in controlling tumor growth may induce autophagic signal pathway.

Raising harmonic variation of arterial pulse in dying rats.

Our previous study revealed that the coefficient of variation of harmonic magnitude (HCV) of radial arterial pulse was significantly raised before the death of cancer patients. In this study, we recorded the caudate arterial pulse of 24 Sprague-Dawley rats that had a fatal dose of urethane injected into their abdomens. Twenty rats were dead within 3 hours after the injection and four survived. We defined the last 100 minutes of each rat's life as the dying process. During the dying process, we found that both the systolic blood pressure and diastolic blood pressure dropped steeply during the last 5 minutes. However, all HCVs, except HCV1, climbed steeply before the last 5 minutes. The HCV1 of the dying rats was significantly higher than that of rats that survived, starting from the first minute (P < 0.01). The HCV2 of the dying rats was significantly higher than that of the survived rats starting from the 52nd minute (P < 0.05). The HCV3 and HCV4 of the dying rats were significantly higher than those of the survived rats until the 70th minute and the 80th minute, respectively (P < 0.05). Furthermore, HCV2-HCV4 proceeded with the dying process and increased gradually. We concluded that HCVs, which failed first in the high-frequency components and then in the low-frequency components, could provide physicians with earlier information to prevent the coming failure of circulatory system, and could reflect quantitatively pathological severity and predict patient outcome. The specific Fourier components in the pulse provide more physiological information than systolic and diastolic blood pressures.

Population structure of wild bananas, Musa balbisiana, in China determined by SSR fingerprinting and cpDNA PCR-RFLP.

Both demographic history and dispersal mechanisms influence the apportionment of genetic diversity among plant populations across geographical regions. In this study, phylogeography and population structure of wild banana, Musa balbisiana, one of the progenitors of cultivated bananas and plantains in China were investigated by an analysis of genetic diversity of simple sequence repeat (SSR) fingerprint markers and cpDNA PCR-RFLP. A chloroplast DNA (cpDNA) genealogy of 21 haplotypes identified two major clades, which correspond to two geographical regions separated by the Beijiang and Xijiang rivers, suggesting a history of vicariance. Significant genetic differentiation was detected among populations with cpDNA markers, a result consistent with limited seed dispersal in wild banana mediated by foraging of rodents. Nuclear SSR data also revealed significant geographical structuring in banana populations. In western China, however, there was no detected phylogeograpahical pattern, possibly due to frequent pollen flow via fruit bats. In contrast, populations east of the Beijiang River and the population of Hainan Island, where long-range soaring pollinators are absent, are genetically distinct. Colonization-extinction processes may have influenced the evolution of Musa populations, which have a metapopulation structure and are connected by migrating individuals. Effective gene flow via pollen, estimated from the nuclear SSR data, is 3.65 times greater than gene flow via seed, estimated from cpDNA data. Chloroplast and nuclear DNAs provide different insights into phylogeographical patterns of wild banana populations and, taken together, can inform conservation practices.

Influence of spleen meridian herbs on the harmonic spectrum of the arterial pulse.

Pulse analysis is a powerful method in Chinese medicine. We suggest that the effect of herbal medicine is to redistribute the blood to organs and meridians. In this report, by injecting extracts into rats and then analyzing the blood pressure wave measured at the caudate arteries, we studied eight important spleen meridian related herbs: They were Semen Lablab, Fructus Amomi Globosi, Rhizoma Atractylodis Macrocephalae, Rhizoma Atractylodis, Tuber Pinelliae, Radix Codonopsitis, Pericarpium Aurantii and Rhizoma Polygonati. All eight herbs increased the intensity of the 3rd harmonic (C3) of the pressure pulse which is correlated to the spleen and spleen meridian, as described in traditional Chinese medical literature. All of them also increased the 2nd harmonic (which is correlated to the kidney and the kidney meridian) as well as decreased the heart load (DC term of pressure wave, C0). Tuber Pinelliae, Radix Codonopsitis, Pericarpium Aurantii and Rhizoma Polygonati decreased the 1st harmonic (correlated to the liver meridian) significantly, while Rhizoma Atractylodis Macrocephalae only decreased C1 slightly. Except for Semen Lablab, all the others decreased the intensity of the 5th (stomach meridian) and the 7th harmonics. The effects of kidney herbs: Cortex Eucommiae and Radix Achyranthis were also shown for comparison.

Liu-wei-dihuang: a study by pulse analysis.

Pulse analysis method was used in studying the traditional Chinese formula Liu-Wei-Dihuang as well as five of its main components (Rehmannia glutinosa, Cornus officinalis, Paeonia Suffruticosa, Poria cocos and Alisma plantogo-aquatica var oriental). Based on our recently developed resonance theory, we tried to elucidate the mechanism and mutual reactions of these meridian related herbs. Hot water herbal extracts were injected intraperitoneally into rats and the pressure pulse spectrum at the caudate artery was measured. The results of this study indicated that Liu-Wei-Dihuang mildly increased blood flow to meridians with lower resonance frequency: namely the liver C1, the kidney C2 and the spleen C3; but decreased blood flow to the higher resonance frequency organs and meridians: the lung C4, the stomach C5, the gall bladder C6, and the bladder C7. It also decreased the heart load C0. All of the five herb components increased blood flow to the kidney C2 and the spleen C3; but their effects on the high frequency organs varied. Alisma plantogo-aquatica var. oriental decreased the C0, C5, C6, C7; Poria cocos decreased C1, C4, C5, C6; Rehmannia glutinosa, Paeonia Suffruticosa decreased C0, C4, C5, C6, C7; Cornus officinalis increased C4 but decreased C0, C5, C6, C7.

Pulse spectrum study on the effect of sie-zie-tang and Radix aconiti.

Extracts of the traditional Chinese formula Sie-Zie-Tang as well as one of its main components, Radix Aconiti were injected into rats intraperitoneally to observe pressure wave spectrum changes at the caudate artery. We found that Radix Aconiti decreased the C0 (DC term of the pulse), C5 and C6 (the harmonic proportions of the 5th and the 6th harmonic), but increased C2 and C3 (the harmonic proportions of the second and the third harmonic) significantly. For Sie-Zie-Tang, the increases of C2, C3, and C4 were accompanied by the decreasing of C0. The decreases of C5, C6 were small and not significant. The additional ingredients in the formula reduce toxic side effects (arrhythmia or heart failure caused by faster and stronger heart beat) due to Radix Aconiti. For human subjects, low dose Sie-Zie-Tang tends to normalize the Fourier components of the pressure wave. Orally taking the formula elevates the harmonic proportion of the harmonic that is lower than normal, but suppresses the higher one. Our results provides a possible mechanism for heart meridian related herbs. It strengthens heart beats, and normalizes energy distribution to different meridians. The study on Sie-Zie-Tang reveals another formula construction to reduce toxic side effects.

Effect of acupuncture at tai-tsih (K-3) on the pulse spectrum.

The frequency specific acupuncture effect on Tai-Tsih (K-3) was examined by investigating pulse variations at the radial artery. The harmonic proportions of the 2nd, 3rd, and the 4th harmonics were increased but the 5th, 6th and 9th harmonics were decreased significantly. The phase angles of all except the 2nd harmonic were increased (wave propagated slower). These results are compared with the frequency specific acupuncture effect on Tsu-San-Li (St-36) which we reported previously (Wang et al., 1995a). This study solidified the weak coupling resonance theory, which successfully explains these frequency specific effects as well as the acupuncture mechanism.

Pulse spectrum analysis of hospital patients with possible liver problems.

Pulse diagnosis were performed on 85 patients who came to the hospital for liver and gall-bladder problems. Correlation between liver tests, which include T-Bil, D-Bil, SGOT, SGPT, ZTT, Alp, gamma-GT, Cho, Alb, and ultra sound scanning, and pulse diagnosis were analyzed. 77 out of 85 subjects showed abnormal liver tests. We used the following 5 criteria for pulse diagnosis as liver abnormality to test the correlation: (1) C1 > or = 3+ and C1 + C4 > or = 4+ or C1 + C6 > or = 4 (in intensity); (2) C1 < or = 3 (in intensity); (3) C6 > or = 3 and C1 + C6 > or = 4 (in intensity); (4) C6 < or = -2 (in intensity) and C6 < or = -2 (in the phase) and (5) C1 > or = 2 C3 < or = -2 (in intensity) or C3 < or = -2 (in the phase). For C1 (liver) every 5% above normal was given one "+," every 5% below normal was given one "-." For C3 (spleen), C4 (lung), C6 (gall-bladder), every 10% above normal was given one "+,", every 10% below normal was given one "-." For the phase, every 10% delay in the traveling speed was given one "-." When considering only the "+" and "-" states and neglecting the quantity of "+" and "-," there are 2(11) (from intensity) x 2(11) (from phase), which equal 2048 x 2048 possible states in the pulse analysis. We considered only 5 criteria for liver abnormality; the correlation was still very high, p < 0.0002, kappa = 0.64. It strongly suggests that meridian theory and pulse diagnosis have physiological and pathological importance.

Pulse spectrum analysis of chemical factory workers with abnormal blood test.

Double blind tests to check the correlation between pulse diagnosis and liver function tests were performed. Blood tests including T-Bil, D-Bil, SGOT and SGPT of 70 chemical factory workers were compared with pulse analysis. Special attention was paid to the indicators of liver and lung meridians for pulse diagnosis. It was found that using the criterion (1) C1 > or = 3+ and C1 + C4 > or = 4+ (hyperfunction) and (2) C1 < or = 3-(hypofunction) as abnormal liver meridian (for C1, every 5% above normal give one +, every 5% below normal give one-; for C4 every 10% above normal give one +, every 10% below normal give one -). The correlation or agreement between the blood tests and the pulse diagnosis was very high (noncorrelation chance checked by X2-test, P < 0.001, degree of agreement checked by Kappa test Ka = 0.61 which means a substantial relationship). Suggestions such as more tests, more criteria and precautions for future study are also proposed.

The prandial effect on the pulse spectrum.

The prandial effect on the pulse spectrum of the radial artery was studied. Ingestion greatly affected the spectrum of the pressure pulse. For most of the tested subjects, the harmonic proportions of the 2nd and the 4th harmonic increased significantly after ingestion, however a decreasing effect was found on the 5th, 6th, 7th, 8th, 9th harmonics. The spectrum became stable from half hour after ingestion and last 3 to 4 hours. From these results, a reliable pulse diagnosis is therefore suggested to be done in the period when the pulse spectrum is stable; physiological significance of these postprandial phenomena is also discussed.

Effect of acupuncture at Tsu San Li (St-36) on the pulse spectrum.

Effect of acupuncture at Tsu San Li (St-36) was examined by investigating the pulse variation of the radial artery. Our results indicated that acupuncture at Tsu San Li has a specific effect on the Fourier components of the pulse. The harmonic proportions were redistributed (C2, C4 decreased, C5, C6, C8 and C9 increased), and the phase angle of the 5th and 8th harmonic waves were decreased (propagating faster). This specific frequency effect was not found when acupuncture needle was applied on a non-acupuncture point. These results can be explained by the resonance theory, which provides a scientific explanation of the acupuncture effect from the hemodynamic view point.

Collective effect of a Chinese formula--a study of xiao-jian-zhong-tang.

Components of a traditional Chinese medicine formula Xiao-Jian-Zhong-Tang were divided into three groups: (1) Radix Paeoniae Lactiflorae, (2) Radix Astragali, and (3) Cinnamon twig, Radix Glycyrrhizae, Fructus jujubae and Saccharum Granorum. Extracts of each group were injected into rats to observe the blood pressure wave spectrum changes at the caudate artery. The whole formula was also extracted and injected into rats to monitor blood pressure wave spectrum changes. Each group has its own effect on the Fourier components of the blood pressure wave. The meridian effects of these herbs are the same as described in the Chinese medical literature, if we consider each meridian in resonance with a specific Fourier component. The whole formula when injected into rats had an effect on the Fourier components of the blood pressure wave similar to the linear combination of individual effect of the three herb groups when used separately. This may be the fundamental idea behind formula construction.

Alteration of pulse in human subjects by three Chinese herbs.

Human subjects were fed with extract of three Chinese herbs, Panax ginseng, Panax quinquefolium roots and Ganoderma lucidum. Pulse of the radial artery was examined. Our results indicate that each herb has a specific effect on the Fourier components of the pulse, and is in agreement with traditional Chinese medical descriptions.

Biomedical engineering basis of traditional Chinese medicine.

Traditional Chinese medicine is an ancient medical science that can be traced back for three to four thousand years of written history. Using the transient response characteristic method, we studied the frequency property of some simulated organs. We tried to explain the concept that the energy in the blood pressure wave is distributed to different organs in the body according to the frequency. This will elucidate the ancient teaching of Chinese medicine in terms of current engineering terminology.

Alterations of pulse by Chinese herb medicine.

Rats were injected with the crude extract of Chinese herbs, Rhizoma Coptidis, Radix Bupleuri and Cinnamomum cassia Blume. The pulse of the tail artery were examined. The results indicated that each drug had a specific effect on the Fourier components of the pulse.