[Current standards for seeds and seedlings of Chinese materia medica and analysis].

Standards for seeds and seedlings of Chinese materia medica are very important to the production, quality control and management of seeds and seedlings of Chinese materia medica. In this paper, the current standards including international standards, national standards, industry standards, local standards and group standards before 2020, involving relevant standards for seeds and seedlings of Chinese materia medica, were comprehensively and systematically summarized. Relevant standards for seeds and seedlings of Chinese materia medica were analyzed based on the standard issue year, the source and types of Chinese materia medica, and whether they are included in the Pharmacopoeia of the People's Republic of China 2015. It is suggested that the standards for seed and seedling of Chinese materia medica shall be systematic, professional and feasible, so as to ensure the sould and sustainable development of the seed and seedling industry of Chinese materia medica.

First evidence of Beauvais' hypothesis in a plant model.

INTRODUCTION: Beauvais presented the application of a so-called 'quantum-like model of homeopathy' by introducing the idea of a type of randomization/unblinding which he called 'in situ'. He predicted that randomized studies based on this type of randomization/unblinding lead to more pronounced effects in placebo controlled randomized homeopathic trials. We designed an experiment regarding wheat germination and stalk length to investigate Beauvais' idea of 'in situ randomization/unblinding' using a homeopathic dilution of sulphur (LM VI) as compared to placebo as well as to water. AIM AND METHOD: The primary aim of this double-blind randomized controlled experiment was to investigate whether there are differences of 'in situ randomization/unblinding' vs 'central randomization/unblinding' with respect to the effect of a homeopathic substance compared to placebo. The secondary aim of our study was to examine possible differences between the sulphur and the placebo group in the 'in situ' arm regarding germination and/or stalk growth of wheat seedlings measured after a seven days exposure. Wheat was treated either with sulphur LM VI, placebo, or water. The wheat grains were placed on glass lids and treatment was performed following the 'in situ randomization/unblinding' as well as 'central randomization/unblinding' method. Germination was measured and classified into three categories. RESULTS: Under 'in situ' randomization/unblinding the odds of a seed not to germinate is 40% lower if treated with sulphur compared to placebo (p=0.004). In contrast, these odds are practically equal in the 'central' meta-group (OR=1.01, p=0.954). Under 'in situ' randomization/unblinding the odds of a seed to germinate with a length ≥1mm is practically equal if treated with sulphur or with placebo (OR=0.96, p=0.717). In contrast, these odds are 21% higher under sulphur compared to placebo in the 'central' meta-group (OR=1.21, p=0.062). In summary, we found a sulphur effect that is significantly different between 'in situ' and 'central' randomization/unblinding relating to all three stages of germination.

Wheat and ultra high diluted gibberellic acid--further experiments and re-analysis of data.

BACKGROUND: Following studies (a) on wheat seedlings and ultra high diluted silver nitrate, and (b) on amphibians and an ultra high diluted hormone, (c) a bio-assay on wheat and extremely diluted gibberellic acid was standardized. This assay was intended to combine the easy-to-handle aspect of (a) and biologically interesting aspects of (b). The purpose of the data analysis presented here was to investigate the influence of an extreme dilution of gibberellic acid on wheat stalk length and to determine the influence of external factors on the experimental outcome. METHODS: Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('G30x'). Analogously prepared water was used for control ('W30x'). 16 experiments including 8000+8000 grains were performed by 9 researchers. RESULTS: Experiments that were performed between January and April showed inconsistent results, whereas most of the experiments performed between September and December showed shorter stalks in the G30x group. This was confirmed by correlation analysis (p<0.01). Thus winter/spring experiments and autumn experiments were analysed separately. When all 10 autumn experiments were pooled, mean stalk lengths (mm) were 48.3±21.4 for the verum group and 52.1±20.4 for control (mean±SD) at grain level (N=5000 per group) and ±5.3 and ±5.1 respectively at dish level. In other words, verum stalk length (92.67%) was 7.33% smaller than control stalk length (100%). The effect size is small when calculation is done on the basis of grains (d=0.18) but, due to the smaller SD at dish level, medium when done on the basis of dishes (d=0.73). The inhibiting effect was observed by 6 of the 6 researchers who performed the autumn experiments. CONCLUSION: The model may be useful for further research as there exists a theoretical justification due to previous studies with wheat and extremely diluted silver nitrate, as well as to previous studies with amphibians and diluted hormones, and its methods are well standardized. Data confirm the hypothesis that information can be stored in the test liquid, even at a dilution of the original substance beyond Avogadro's value; and that the wheat bio-assay is sensitive to such information.

Wheat and ultra high diluted silver nitrate--further experiments and re-analysis of data.

BACKGROUND: Since 1926, an influence of a dilution of silver nitrate (24x) on the growth of coleoptiles of wheat seedlings was described. The aim of the study discussed here is the critical proof of the reliability of a test system which has been quoted as a basic model for the research on homoeopathy for decades. METHODS: Grains of winter wheat (Triticum aestivum) were observed under the influence of extremely diluted silver nitrate (10(-23)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('24x'). Analogously prepared water and/or inert water was used for control. Thirty experiments including 5000+5000 grains were performed by 5 researchers. RESULTS: Stalk lengths clearly indicate that development is enhanced by the probe silver nitrate 24x as compared to control. When the experiments 1989-1995 were pooled, means and SD for silver nitrate 24x-groups were 42.3±26.9 mm and for water control groups 34.7±22.2 mm. Verum stalk length was 21.9% bigger than control (100%) (p<0.01; d=0.31, i.e. small). For the experiments 1998-2014, means and SD were 73.7±21.7 mm and 60.5±20.9 mm. Verum stalk length was 21.7% bigger than control (100%) (p<0.01; d=0.62, i.e. medium). From the results one may hypothesize that the result is more marked in experiments showing an average mean of stalk length between ca. 50 and 90 mm in contrast to smaller or bigger mean lengths. CONCLUSION: The previous findings were confirmed by the results.

Stimulative influence of germination and growth of maize seedlings originating from aged seeds by 2,4-D potencies.

BACKGROUND: The 2,4-D (2,4-dichlorophenoxyacetic acid) is using as a growth regulator in tissue culture media. Maize seeds have poor ability to maintain germination rate in the long term. OBJECTIVE: To examine the possible restorative effect of homeopathic 2,4-D potencies on maize seedlings originating from seeds damaged by accelerated aging. METHODS: Seeds of four maize lines were subjected to accelerated aging stress treatment. Seed samples were treated with distilled water (control) and a range of potencies of 2,4-D: 3C, 3.75C, 4.5C, 5.25C and 6C. The germination capacity, fresh substance (FS) and length of root and shoot were determined. Hydrolysis and biosynthesis, GSH/GSSG ratio and redox capacity were calculated. RESULTS: Induced seed aging decreased germination rate and growth of seedlings. 2,4-D potencies did not have a statistically significant effect on germination. However, there were statistically significant effects on FS production, root and shoot length and redox capacity. The 3C potency had the largest effect on the FS accumulation, 4.5C increased root and shoot length, compared to control (statistically significant). The GSH/GSSG ratio and the redox capacity were decreased by aging. The 3C and 4.5C potencies tended to reverse the GSH/GSSG ratio (statistically significant) in the root and shoot, (i.e., shifted the redox balance to the reduced state). CONCLUSION: Homeopathic potencies of 2,4-D appear to have a beneficial effect on artificially aged maize seeds: they stimulate growth through better substance conversion from seed rest, and shift the redox capacity towards a reduced environment. Further work is required to determine if this is an useful means of improving maize seed germination and growth.

Zincum Metallicum, a homeopathic drug, alleviates Zn-induced toxic effects and promotes plant growth and antioxidant capacity in Lepidium sativum L.

In this study, we investigated the effect of the homeopathic drug Zincum Metallicum (ZM) on zinc (Zn) toxicity in the plant species Lepidium sativum L. We focused on growth parameters, Zn uptake and numerous biochemical parameters. Seedlings were hydroponically subjected during 7 days to 0.05, 500, 1000, 1500 and 2000 µM Zn2+, in the absence or presence of 15ch or 9ch ZM. In the absence of ZM, Zn induced negative effect on growth especially at the dose of 2 mM. Zn induced also chlorosis, reduced total chlorophyll and/or carotenoid content and increased the level of malondialdehyde (MDA). Under Zn toxicity (500, 1000 and 1500 µM), the superoxide dismutase (SOD), catalase (CAT), gaiacol peroxidase (GPX) and glutathione reductase (GR) activities were increased or not significantly affected, while at 2000 µM Zn affected the activity of these enzymes. At the highest Zn level (2 mM), proline and total polyphenol and flavonoid contents were markedly increased in leaves and roots of L. sativum. Additionally, ZM supply considerably ameliorated the plant growth, photosynthetic pigment contents and increased non-enzymatic antioxidant molecules and enzymatic activities against Zn-induced oxidative stress. Our data suggest that homeopathic properties of ZM may be efficiently involved in the restriction of Zn-induced oxidative damages, by lowering Zn accumulation and translocation in the leaves and roots of Lepidium sativum L.

Homeopathic preparations to control the rosy apple aphid (Dysaphis plantaginea Pass.).

A laboratory model system with the rosy apple aphid (Dysaphis plantaginea Pass.) on apple seedlings was developed to study the effects of homeopathic preparations on this apple pest. The assessment included the substance Lycopodium clavatum and a nosode of the rosy apple aphid. Each preparation was applied on the substrate surface as aqueous solution of granules (6c, 15c, or 30c). Controls were aqueous solutions of placebo granules or pure water. In eight independent, randomized, and blinded experiments under standardized conditions in growth chambers, the development of aphids on treated and untreated apple seedlings was observed over 17 days, each. Six experiments were determined to assess the effects of a strict therapeutic treatment; two experiments were designed to determine the effects of a combined preventative and therapeutic treatment. After application of the preparations, the number of juvenile offspring and the damage on apple seedlings were assessed after 7 and 17 days, respectively. In addition, after 17 days, the seedling weight was measured. In the final evaluation of the six strictly therapeutic trials after 17 days, the number of juvenile offspring was reduced after application of L. clavatum 15c (-17%, p = 0.002) and nosode 6c (-14%, p = 0.02) compared to the pure water control. No significant effects were observed for leaf damage or fresh weight for any application. In the two experiments with combined preventative and therapeutic treatment, no significant effects were observed in any measured parameter. Homeopathic remedies may be effective in plant-pest systems. The magnitude of observed effects seems to be larger than in models with healthy plants, which renders plant-pest systems promising candidates for homeopathic basic research. For successful application in agriculture, however, the effect is not yet sufficient. This calls for further optimization concerning homeopathic remedy selection, potency level, dosage, and application routes.

A review of three simple plant models and corresponding statistical tools for basic research in homeopathy.

In this paper, we review three simple plant models (wheat seed germination, wheat seedling growth, and infected tobacco plants) that we set up during a series of experiments carried out from 1991 to 2009 in order to study the effects of homeopathic treatments. We will also describe the set of statistical tools applied in the different models. The homeopathic treatment used in our experiments was arsenic trioxide (As2O3) diluted in a decimal scale and dynamized. Since the most significant results were achieved with the 45th decimal potency, both for As2O3 (As 45x) and water (W 45x), we here report a brief summary of these results. The statistical analysis was performed by using parametric and nonparametric tests, and Poisson distribution had an essential role when dealing with germination experiments. Finally, we will describe some results related to the changes in variability, which seems to be one of the targets of homeopathic treatment effect.

Isopathic treatment effects of Arsenicum album 45x on wheat seedling growth--further reproduction trials.

BACKGROUND: Two experimental studies on wheat preintoxicated with Arsenic trioxide yielded a significant shoot growth increase after an isopathic application of Ars-alb 45x. One independent reproduction trial however, yielded an effect inversion: wheat shoot growth was significantly decreased after application of Ars-alb 45x. AIMS: In this study we investigated the role of three potential confounding factors on the experimental outcome: geographical location of the experiments, influence of the main experimenter, and seed sensitivity to Arsenic poisoning. Laboratory-internal reproducibility was assessed by meta-analysis. MATERIAL AND METHODS: Wheat poisoned with Arsenic trioxide was cultivated in vitro in either Ars-alb 45x, water 45x, or unpotentised water. Treatments were blinded and randomised. Shoot length was measured after 7 days. The stability of the experimental set-up was assessed by systematic negative control (SNC) experiments. RESULTS: The SNC experiments did not yield significant differences between the three groups treated with unpotentised water. Thus the experimental set-up seemed to be stable. We did not observe any shoot growth increase after a treatment with Ars-alb 45x in any of the newly performed experiments. In contrast, the meta-analysis of all 17 experiments performed (including earlier experiments already published) yielded a statistically significant shoot growth decrease (-3.2%, p=0.017) with isopathic Ars-alb 45x treatment. This effect was quantitatively similar across all five series of experiments. CONCLUSIONS: Ultramolecular Ars-alb 45x led to statistically significant specific effects in arsenic poisoned wheat when investigated by two independent working groups. Effect size and effect direction differ, however. The investigated factors (geographical location, experimenter, seed sensitivity to Arsenic poisoning) did not seem to be responsible for the effect inversion. Laboratory external reproducibility of basic research into homeopathic potentisation remains a difficult issue.

The influence of ultra-high diluted compounds on the growth and the metabolites of Oryza sativa L

The ultra-high dilutions (UHDs) have been widely used in the field of human, animal and plant treatment. In the present research, the effects of the potentized ultra-high dilutions (UHDs) on physiological and biochemical variations in Oryza sativaL. (rice) were investigated. Methods: To study the effect of UHDs (Calendula officinalisCalen. and Arnica montanaArn.) on the physiological and biochemical variations of the Oryza sativaL. (rice), 28 experiments were designed and statistically analyzed using the Design Expert 7.0.1 software over the general factorial design methodology. Three qualitative factors were studied including the UHDs/placebo usage, sterile/non-sterile experimental condition and the type and timing of the UHDs usage. The validated analysis was subjected to more extended studies on the variations in physiological growth, carbohydrate, protein content, pigment production, and amino acid patterns. To evaluate the effects of UHDs on rice, a desirable response percentage was formed from a number of healthy seedling productions, and the height percentage of the aerial parts and main roots were studied. Results: The statistical analysis resulted in a prediction model which was more than 97% correlates with experimental results. The results showed that the UHDs increased the pH variations, carbohydrate, protein and pigment levels each by ~2.5, ~1.5, ~1.4, and ~1.4 folds, respectively. Also compared to placebo, the amount and proportionof amino acids has significantly varied, showing a statistical effect on the germination and seedling growth of the rice, as well as the stress conditions caused by the sterilization process, seedlings entrance into the light and their transition into hydroponic culture medium.Conclusion: The use of UHDs leads to an increase in the production of chlorophyll, as well as carbohydrate and protein content. Moreover, it causes significant variations in the amino acid profile and the production of amino acids along with the photosynthesis, germination, and metabolism processes.(AU)

The effect of extremely diluted agitated gibberellic acid (10e-30) on wheat stalk growth--a two researcher pilot study.

OBJECTIVE: Use of a wheat growth bio assay after 7 days in research on homeopathic dilutions of gibberellic acid. METHODS: Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy (30×). Analogously prepared water was used for control. In a two centre study, 3 experiments with a total of 4880 grains were performed. RESULTS: Data were found to be rather homogeneous within the control group as well as within the verum group in general. Germination rates were around 95%, with no significant difference between verum and control group (p>0.05). Mean stalk lengths (mm) were 40.63±20.96 for the verum and 44.33±21.11 for the control group (mean±S.D.) at grain level (N=2440 per group) and ±5.33 and ±5.89, respectively at dish level (122 cohorts of 20 grains per treatment group). In other words, verum stalk length (91.65%) was 8.35% smaller than control stalk length (100%). This difference is statistically highly significant (p<0.001) and was found by both researchers involved independently. CONCLUSION: These results suggest that there was an influence of gibberellic acid 30× on wheat seedling development, i.e. the wheat growth bio assay can be a useful tool for further experiments on homeopathic dilutions of gibberellic acid.

Sepia 200cH at 1: 1000 dilution ameliorates salt stress in cowpea seedlings but its medium 90 por cento ethanol proves ineffective at the same dilution

Soil salinity severely affects crop yield all over the world. In a recent study we observed that Natrum mur 200cH, a homeopathic remedy, improved growth in germinating cowpea seeds. In the present study we have tested another remedy Sepia, which is complementary to Natrum mur, on cowpea seedlings under salt stress. Cowpea seedlings grown over moist filter paper in petridishes were divided into 4 groups: (1) control in sterile water, (2) in 50mM NaCl solution, (3) seeds pretreated with 90% ethanol diluted with water 1:100 and then transferred to 50mM NaCl solution, (4) seeds pretreated with Sepia 200cH diluted with water 1:100 and transferred to 50mM NaCl solution. In another experiment the groups were same, but the dilution of 90% ethanol and Sepia 200cH was 1:1000 instead of 1:100. The purpose was to further reduce the ethanol content in both the drug and its vehicle 90% ethanol, so that the alcohol effect is minimized or abolished. The data were analysed by ANOVA followed by t-test. Sepia 200cH at both 1:100 and 1:1000 dilutions significantly increased growth, sugar, chlorophyll, protein and water content in seedlings as compared to the untreated salt-stressed group. The effect with the1000th dilution of Sepia 200cH was more pronounced than with its 100th dilution. The vehicle 90% ethanol at 1:100 dilution produced some positive effect on the seedlings, but the 1000th dilution of the vehicle produced no such effect. It is, therefore, concluded that Sepia 200cH could ameliorate salt stress in cowpea seedlings and that the 1000th dilution is more effective than its 100th dilution. The alcohol effect is totally eliminated with the 1000th dilution of 90% ethanol. Thus the 1000th dilution could retain the drug effect and eliminate the vehicle effect.

Introdução: A salinidade do solo afeta seriamente a produtividade das culturas no mundo todo. Num estudo recente, observamos que o medicamento homeopático Natrum muriaticum 200 Ch melhorou o crescimento de sementes de feijão-fradinho Vigna unguiculata (L.) Walp] em germinação. Objetivo: Nesse trabalho, testamos o medicamento homeopático Sepia succus, complementar de Nat-m, em plântulas de feijão-fradinho submetidas a estresse salino. Métodos: Plântulas de feijão-fradinho desenvolvidas sobre papel de filtro umedecido em placas de Petri foram divididas em 4 grupos: (1) controle em água estéril, (2) em solução de NaCl 50 mM, (3) sementes pré-tratadas com etanol a 90% diluído 1:100 em água e após transferido a solução de NaCl 50mM, e (4) sementes pré-tratadas com Sep 200 cH diluídas 1:100 em água e após transferidas a solução de NaCl 50 mM. Num segundo experimento, os grupos foram os mesmos, mas a diluição do etanol a 90% e de Sep 200 cH foi 1:1.000 ao invés de 1:00 para reduzir ainda mais o teor de etanol em ambos, medicamento e veículo a fim de minimizar ou abolir o efeito do álcool. Os dados foram analisados através de ANOVA seguida de teste t de Student. Resultados: As duas diluições de Sep 200 cH, 1:100 e 1:1.000, aumentaram significativamente o crescimento, teor de açúcar, clorofila, proteína e água das plântulas por comparação ao grupo submetido a estresse salino não tratado. O efeito da 1.000ª diluição de Sep 200 cH foi mais pronunciado que o da 100ª diluição. O veículo etanol a 90% em diluição 1:100 induziu alguns efeitos positivos nas plântulas, mas a diluição 1:1.000 não apresentou esses efeitos. Conclusões: Sep 200 cH reverteu o efeito de estresse salino nas plântulas de feijão-fradinho, sendo que a 1.000ª diluição foi mais efetiva que a 100ª. O efeito do álcool foi totalmente eliminado com a 1.000ª diluição do etanol a 90%. Portanto, a 1.000ª conservou o efeito do medicamento e eliminou o efeito do veículo.

Sepia 200cH at 1: 1000 dilution ameliorates salt stress in cowpea seedlings but its medium 90% ethanol proves ineffective at the same dilution

Soil salinity severely affects crop yield all over the world. In a recent study we observed that Natrum mur 200cH, a homeopathic remedy, improved growth in germinating cowpea seeds. In the present study we have tested another remedy Sepia, which is complementary to Natrum mur, on cowpea seedlings under salt stress. Cowpea seedlings grown over moist filter paper in petridishes were divided into 4 groups: (1) control in sterile water, (2) in 50mM NaCl solution, (3) seeds pretreated with 90% ethanol diluted with water 1:100 and then transferred to 50mM NaCl solution, (4) seeds pretreated with Sepia 200cH diluted with water 1:100 and transferred to 50mM NaCl solution. In another experiment the groups were same, but the dilution of 90% ethanol and Sepia 200cH was 1:1000 instead of 1:100. The purpose was to further reduce the ethanol content in both the drug and its vehicle 90% ethanol, so that the alcohol effect is minimized or abolished. The data were analysed by ANOVA followed by t-test. Sepia 200cH at both 1:100 and 1:1000 dilutions significantly increased growth, sugar, chlorophyll, protein and water content in seedlings as compared to the untreated salt-stressed group. The effect with the1000th dilution of Sepia 200cH was more pronounced than with its 100th dilution. The vehicle 90% ethanol at 1:100 dilution produced some positive effect on the seedlings, but the 1000th dilution of the vehicle produced no such effect. It is, therefore, concluded that Sepia 200cH could ameliorate salt stress in cowpea seedlings and that the 1000th dilution is more effective than its 100th dilution. The alcohol effect is totally eliminated with the 1000th dilution of 90% ethanol. Thus the 1000th dilution could retain the drug effect and eliminate the vehicle effect. (AU)

Introdução: A salinidade do solo afeta seriamente a produtividade das culturas no mundo todo. Num estudo recente, observamos que o medicamento homeopático Natrum muriaticum 200 Ch melhorou o crescimento de sementes de feijão-fradinho Vigna unguiculata (L.) Walp] em germinação. Objetivo: Nesse trabalho, testamos o medicamento homeopático Sepia succus, complementar de Nat-m, em plântulas de feijão-fradinho submetidas a estresse salino. Métodos: Plântulas de feijão-fradinho desenvolvidas sobre papel de filtro umedecido em placas de Petri foram divididas em 4 grupos: (1) controle em água estéril, (2) em solução de NaCl 50 mM, (3) sementes pré-tratadas com etanol a 90% diluído 1:100 em água e após transferido a solução de NaCl 50mM, e (4) sementes pré-tratadas com Sep 200 cH diluídas 1:100 em água e após transferidas a solução de NaCl 50 mM. Num segundo experimento, os grupos foram os mesmos, mas a diluição do etanol a 90% e de Sep 200 cH foi 1:1.000 ao invés de 1:00 para reduzir ainda mais o teor de etanol em ambos, medicamento e veículo a fim de minimizar ou abolir o efeito do álcool. Os dados foram analisados através de ANOVA seguida de teste t de Student. Resultados: As duas diluições de Sep 200 cH, 1:100 e 1:1.000, aumentaram significativamente o crescimento, teor de açúcar, clorofila, proteína e água das plântulas por comparação ao grupo submetido a estresse salino não tratado. O efeito da 1.000ª diluição de Sep 200 cH foi mais pronunciado que o da 100ª diluição. O veículo etanol a 90% em diluição 1:100 induziu alguns efeitos positivos nas plântulas, mas a diluição 1:1.000 não apresentou esses efeitos. Conclusões: Sep 200 cH reverteu o efeito de estresse salino nas plântulas de feijão-fradinho, sendo que a 1.000ª diluição foi mais efetiva que a 100ª. O efeito do álcool foi totalmente eliminado com a 1.000ª diluição do etanol a 90%. Portanto, a 1.000ª conservou o efeito do medicamento e eliminou o efeito do veículo. (AU)