Efficacy of the herbal pair, Radix Achyranthis Bidentatae and Eucommiae Cortex, in preventing glucocorticoid-induced osteoporosis in the zebrafish model.

OBJECTIVE: In traditional Chinese medicine, the herbal pair, Radix Achyranthis Bidentatae (RAB) and Eucommiae Cortex (EC), is widely used to treat osteoporosis. Herein, we determined whether this herbal pair can be used to ameliorate glucocorticoid (GC)-induced osteoporosis (GIOP) and find its optimal dosage in zebrafish. METHODS: The characteristics of the aqueous extract of RAB and EC were separately characterized using high-performance liquid chromatography. Osteoporosis was induced in 5-day post-fertilization zebrafish larvae by exposing them to 10 µmol/L dexamethasone (Dex) for 96 h. Seven combinations of different ratios of RAB and EC were co-administered. Treatment efficacy was determined by calculating zebrafish vertebral area and sum brightness, via alizarin red staining, and by detecting alkaline phosphatase (ALP) activity. Multiple regression analysis was conducted to test the optimal dosage ratio. RESULTS: According to the Chinese Pharmacopoeia (2015), ß-ecdysone (ß-Ecd) is a major bioactive marker in RAB extract, while pinoresinol diglucoside (PDG) is the major marker in EC extract. Both of ß-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards. Treatment with 10 µmol/L Dex reduced zebrafish vertebral area, sum brightness, and ALP activity, but RAB and EC attenuated these effects. Combining 50 µg/mL RAB and 50 µg/mL EC was optimal for preventing GIOP in zebrafish. Reverse transcription-quantitative polymerase chain reaction was used to evaluate the mRNA expression of osteogenesis-related genes. A treatment of 10 µmol/L Dex decreased runt-related transcription factor 2 (Runx2), osteogenic protein-1 (OP-1), bone γ-carboxyglutamic acid-containing protein (BGLAP), and ß-catenin levels. This effect was counteracted by RAB and EC co-treatment (P < 0.05). Additionally, the effect of using the two herbal extracts together was better than single-herb treatments separately. These results demonstrated that RAB and EC preserve osteoblast function in the presence of GC. The best mass ratio was 1:1. CONCLUSION: RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish, with 1:1 as the optimal dosage ratio.

Beta-Ecdysone Protects Mouse Osteoblasts from Glucocorticoid-Induced Apoptosis In Vitro.

Glucocorticoid-induced osteoporosis is a common form of secondary osteoporosis. Glucocorticoids affect both bone formation and resorption, and prolonged glucocorticoid exposure can suppress osteoblast activities. beta-Ecdysone, found in many plants, is involved in protein synthesis, carbohydrate and lipid metabolism, and immunologic modulation. Here, we evaluated the effects of beta-ecdysone on osteoblast viability by assessing apoptosis following treatment with excess glucocorticoids. Mouse bone marrow stromal cells were induced to differentiate and grow into osteoblasts, and then treated with 10 µM glucocorticoid and 10, 1, or 0.1 µM beta-ecdysone. The expression levels of osteoblast growth and differentiation factors (runt-related transcription factor 2, osteogenic protein-1, and alkaline phosphatase), apoptosis-related genes (transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8), and Akt1 and phospho-Akt (Thr308) were then assessed via alkaline phosphatase staining, acridine orange-propidium iodide staining, annexin V/PI apoptosis assay, real-time RT-PCR, and Western blot analyses. Notably, treatment with 10 µM glucocorticoid resulted in reduced osteoblast viability and the specific activity of alkaline phosphatase as well as reduced runt-related transcription factor 2, osteogenic protein-1, and alkaline phosphatase mRNA expression in vitro, indicating that glucocorticoid inhibited osteogenic differentiation. Moreover, glucocorticoid treatment yielded increased transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8 expression and decreased Akt1 and phospho-Akt levels, indicating glucocorticoid-induced apoptosis. Meanwhile, beta-ecdysone inhibited glucocorticoid function, preserving the expression of Akt1 and phospho-Akt and reducing the expression of transformation-related protein 53, ataxia telangiectasia mutated protein, caspase-3, and caspase-8. Thus, beta-ecdysone prevented glucocorticoid-induced osteoblast apoptosis in vitro. These data highlight the potential for beta-ecdysone as a treatment for preventing the effects of glucocorticoid on bone growth.

Shenfu Injection suppresses inflammation by targeting haptoglobin and pentraxin 3 in rats with chronic ischemic heart failure.

OBJECTIVE: To investigate the regulatory effects of Shenfu Injection (SFI, ) on hemodynamic parameters and serum proteins in rats with post-infarction chronic heart failure (CHF). METHODS: Forty-five healthy Wistar rats were randomized into three groups: sham, heart failure (model) and SFI group. The CHF was induced by left coronary artery ligation. Seven days after the surgical operation, animals in the sham group and the model group received saline (6.2 mL/kg/d), while animals in the SFI group received SFI (6.2 mL/kg d) intraperitoneally. Four weeks later, cardiac hemodynamic parameters were measured via the carotid route. The expression of serum proteins was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS). RESULTS: Recording of hemodynamic parameters showed that left ventricular systolic pressure (LVSP), maximum rate of left ventricular pressure (+dp/dtmax) rise, and maximum rate of left ventricular pressure (-dp/dtmax) decrease, while the left ventricular end diastolic pressure (LVEDP) rose in the model group compared to those in the sham group (P <0.05). The results of the MALDI-TOF MS indicated that haptoglobin (HP), pentraxin 3 (PTX3) and alpha-1-antitrypsin were up-regulated, while serum albumin and 40S ribosomal protein were down-regulated in the model group (P <0.05). Compared with the model group, LVSP, +dp/dtmax and -dp/dtmax were higher, while LVEDP was lower in the SFI group (P<0.05). Expression levels of HP and PTX3 were lower than in the model group (P<0.05). CONCLUSION: SFI could improve hemodynamic function and decrease inflammatory reactions in the pathophysiology of CHF. The serum proteins HP and PTX3 could be potential biomarkers for chronic ischemic heart failure, and they could also be the serum protein targets of SFI.

[Effects of glycyrrhetinic acid on sodium ion channel currents of rats' ventricular myocardial cells].

OBJECTIVE: To study the effects of glycyrrhetinic acid (GA) on the sodium ion channel currents (I(Na)) of rats' ventricular myocardial cells, and to explore its anti-arrhythmic mechanisms at the ion channel level. METHODS: Single ventricular myocardial cells was isolated from SD rats. The whole cell patch clamp was used to record the effects of GA on I(Na) of rats' ventricular myocardial cells. RESULTS: GA could inhibit I(Na) of rats' ventricular myocardial cells dose-dependently. GA at 1, 5, and 10 micromol/L decreased I(Na) of rats' ventricular myocardial cells from (-4.26 +/- 0.15) nA to (-3.54 +/- 0.10) nA, (-2.19 +/- 0.09) nA, and (-1.25 +/- 0.08) nA, respectively. GA at 1, 5, and 10 micromol/L inhibited I(Na) by 16.08% +/- 2.3%, 50.82% +/- 3.56%, and 75.98% +/- 5.12%, showing statistical difference when compared with the control group (P < 0.05). GA at 10 micromol/L shifted I(Na) current-voltage curve more positively, but the activation potential and the peak potential were not changed. CONCLUSION: GA inhibited the I(Na) of rats' ventricular myocardial cells dose-dependently, which was possibly associated with its antiarrhythmia effects.

[Analysis on pulse diagram characteristics of subjects with subhealth state].

OBJECTIVE: To study the pulse diagram parameters of subjects with subhealth state and to find the pulse parameters for subhealth state evaluation. METHODS: A total of 1 275 subjects without diagnosed diseases were recruited and their health conditions were assessed with Health Evaluating Questionnaire H20 V2009. The subjects were assigned to health group or subhealth group according to the scale score. Subjects' syndrome in the subhealth group was differentiated using score of "subhealth state of syndrome differentiation V2010". Another 121 patients with cardiovascular diseases were enrolled as a control. The pulse information was collected with YJJ-101 subhealth pulse monitoring system and the parameters include amplitude of main wave (h1), amplitude of repeat wave (h5) and its front wave (h3), 1/3 or 1/5 width of main wave (w1) or (w2), time of rapid ejection phase (t2), period of pulse (t), pulse pressure (Pp), square (S), area in systole (As) and area in diastole (Ad) of pulse diagram and ratios of h3/h1, h5/h1, w1/t, w2/t and h1/t1. RESULTS: Pulse diagram analysis showed significant differences among health, subhealth and disease group in Pp, h1, S and As and ratios of h5/h1 and w2/t. Compared with the health group, the values of w1/t and w2/t of the subhealth group increased (P<0.05), and Pp, h1, h5, h5/h1, S, As and Ad decreased (P<0.05). Compared with health group, the parameters of pulse of the subhealth group were increased in Pp and h5/h1 (P<0.05) and decreased in h1, w2/t, S and As (P<0.05). Compared with health group, pulse parameters h3/h1, w1, w1/t, w2/t of excess and deficiency syndrome group increased, and h1, h5, h1/t 1and h5/h1 decreased. Among different syndromes of subhealth state, pulse diagram parameters h1, h5, h3/h1, h5/h1 and w1/t of yin deficiency, qi deficiency, liver stagnation and excess heat group were significantly different (P<0.05) from the health group, for example, pulse parameters h1 and h5 of stagnation, yin deficiency, qi deficiency and excess heat group declined in order, and pulse parameters h3/h1 and w1/t of liver stagnation, excess heat, yin deficiency and qi deficiency group increased in order. Pulse index h1 in the kidney deficiency group was higher than that in the health group and the other syndrome groups. CONCLUSION: Results of analyzing sphygmogram parameters showed different characteristics among different health status and the subhealth state due to different syndromes. Sphygmogram parameters may be used for objective evaluation of health status or subhealth syndrome differentiation.

Roles and mechanisms of ginseng in protecting heart.

Ginseng, the root of Panax ginseng C. A. Mayer, has long been used clinically in China to treat various diseases. Multiple effects of ginseng, such as antitumor, antiinflammatory, antiallergic, antioxidative, antidiabetic and antihypertensive have been confirmed by modern medicine. Recently, the clinical utilization of ginseng to treat heart diseases has increased dramatically. The roles of ginseng in protecting heart are foci for research in modern medical science and have been partially demonstrated, and the mechanisms of protection against coronary artery disease, cardiac hypertrophy, heart failure, cardiac energy metabolism, cardiac contractility, and arrhythmia are being uncovered progressively. However, more studies are needed to elucidate the complex mechanisms by which ginseng protects heart. All such studies will provide evidence of ginseng's clinical application, international promotion, and new drug development.

[Facial spectrum and color characteristics of five viscera reflecting on the face in sub-health status].

OBJECTIVE: To observe the facial spectrum and color of different points, the positions of heart, liver, spleen, lung and kidney of traditional Chinese medicine reflecting on the face, in healthy participants and those with a sub-health status, so as to provide an objective basis for health evaluation. METHODS: The health condition of 470 subjects without acute and chronic conditions was assessed using the Health Evaluating Questionnaire H20 V2009. The subjects were diagnosed with health (more than 80 score) or sub-health (score between 60 and 80) status according to the questionnaire score. The subjects with a subhealth status were also analyzed using the five-viscera syndrome differentiation of traditional Chinese medicine using the form for collecting information according to the four examinations. Then for gathering the facial color information, CIE L*a*b and C values and reflections of wavelengths ranged from 400 to 700 nm were measured using a CM-2600D spectrophotometer on 8 points of the face, including the frontal part, glabella, nose, mandible, two cheeks and eyelids. RESULTS: L value of the sub-health group was higher than that of the health group (P<0.05), and a, b and C values were lower than those of the health group (P<0.05), suggesting that the facial complexion of the sub-health group was pale/whiter than the health group. The reflectance rates of wavelengths (from 400 to 550 nm) of the sub-health group were higher than those of the health group (P<0.05), which suggested that the facial complexion of the sub-health group was partially green. The a, b and C values of the forehead, glabella and nose of subjects in the sub-health group were apparently different from the health subjects. L values of five-viscera types were significantly different from people of a health status (P<0.05). There were some differences in color values among the five-viscera groups: lung group's color values were significantly different from the others in terms of a, b and C values. The spectral reflectance of different viscera groups of the sub-health group also showed certain differences: reflectance of wavelengths at 400 to 490 nm in the lung group and at 520 to 580 nm in the spleen group showed significant difference from the other groups, and the lung group was lower and the spleen group was higher than the others. CONCLUSION: There are some differences in facial spectrum and color in different sites of the face in sub-health status of different viscera syndrome types, which can provide an objective basis for health evaluation.

Effect of matrine on human ether à go-go related gene (HERG) channels expressed in Chinese hamster ovary cells.

OBJECTIVE: To observe the effect of matrine on human ether à go-go related gene (HERG) potassium channels expressed in Chinese hamster ovary (CHO) cells and investigate whether HERG channel is a new target of the pharmacological effect of matrine on arrhythmia and tumor METHODS: HERG channel potassium current in CHO cell was recorded using whole-cell patch-clamp technique, and the influence of matrine on the current was explored. RESULTS: Matrine inhibited HERG potassium current in a dose-dependent manner, and the 50% inhibitory concentration (IC IC(50)) was 411±23 µmol/L. Matrine had no significant effect on the activation kinetics, and mainly blocked HERG channels in their closed state. CONCLUSIONS: The blocking effect of matrine on HERG channels might be one of the mechanisms against arrythmias and tumors. Unlike most other blockers exerting blocking effect at the intracellular sites by entering the cell with the opening of HERG channel, matrine blocked HERG channels at the extracellular sites.

[Effects of fufangdenshen tablets on human atherosclerotic plasma protein using proteomic technique].

OBJECTIVE: To research the effects of fufangdenshen tablets on atherosclerotic plasma protein using proteomic technique and further explore its potential molecular mechanism to cure atherosclerosis. METHOD: The plasma of normal, atherosclerosis and cured group were collected, and the albumin in plasma was removed. Proteomic protocol involved of 2-DE, image analysis and spectrometry detection was used to detect regulated plasma protein by fufangdenshen tablets. RESULT: 3 decreased expressed and 6 higher protein in atherosclerotic plasma could be recovered by fufangdenshen tablets. The levels of fibrinogen, Granzyme C and immunoglobulin were decreased by fufangdenshen tablets. CONCLUSION: The levels of fibrinogen, Granzyme C in atherosclerotic plasma could be decreased by fufangdenshen tablets. The effects of fufangdenshen tablets on atherosclerosis included that: inhibition of adhesion of monocyte, inhibition of proliferation and migration of VSMC, and weakening of inflammation.

[Effect of electroacupuncture at "Neiguan" (PC 6) on expression of c-fos gene in myocardium of the rat with acute myocardial ischemia].

OBJECTIVE: To explore the mechanism of electroacupuncture at "Neiguan" (PC 6) improving acute myocardial ischemia. METHODS: The rats were randomly divided into a sham operation group, a myocardial ischemia model group and a myocardial ischemia model plus electroacupuncture group. The acute myocardial ischemia model was developed byligation of the descending anterior branch of the coronary artery, and electroacupuncture was given at bilateral "Neiguan" (PC 6). Serum myocardial enzymes was determined by biochemical method and the expression of c-fos mRNA in myocardium was detected by using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The activities of serum myocardial enzymes and the expression of c-fos mRNA in ischemic myocardium were significantly increased as compared with those in the sham operation group (P < 0.05), and after electroacupuncture they were significantly decreased (P < 0.05). CONCLUSION: The mechanism of electroacupuncture at "Neiguan" (PC 6) improving acute myochadial ischemia is possibly related with down-regulation of expression of c-fos mRNA in myocardium.

[Clinical observation on treatment of 40 SARS uncertain patients with integrative traditional Chinese and Western medicine].

OBJECTIVE: To observe the clinic symptom improving time in uncertain SARS patients and the therapeutic effect of integrative Chinese and western medicine (ICWM) in treating SARS. METHODS: The clinic symptoms, chest film and tongue figure of 40 uncertain SARS patients treated with ICWM were observed and T-lymphocyte subsets, serum coronavirus nucleic acid and antibody in 20 patients were tested dynamically. RESULTS: All the symptoms, such as fever, sweating, fatigue, cough without phlegm, etc. were obviously improved after treatment. Lung shadow in chest film began to be absorbed 4.54 +/- 2.85 days, and obviously absorbed 7.74 +/- 4.68 days after treatment. CD3, CD4 and CD8 in 20 patients, which were lower than the normal range when hospitalization, began to increase 3 days later and gradually recovered to the normal in 6-10 days. Serum coronavirus nucleic acid was positive in 3 patients, coronavirus antibody positive in two and both were positive in one. CONCLUSION: ICWM can improve the symptoms and regulate the immune function in uncertain SARS patients.