[Identification of Two GLOBOSA-like MADS-box Genes in Tea Plant (Camellia sinensis [L.] O. Kuntze)].

Tea plant (Camellia sinensis [L.] O. Kuntze) is a woody crop of high economic importance worldwide; however, information on the molecular mechanisms underlying the regulation of flower development in this species is limited. In the present study, two GLOBOSA (GLO) -like MADS-box genes, CsGLO1 and CsGLO2, were isolated from C. sinensis 'Ziyangzhong' and were characterized to elucidate their roles in flower development. We found that CsGLOl and CsGLO2 are nuclear-localized transcription factors without transactivation ability but with a robust interaction. They have similar patterns of expression, both mainly restricted to petals and stamens. Moreover, ectopic expression of either CsGLO1 or CsGLO2 in Arabidopsis thaliana resulted in a partial conversion of sepals to petals, suggesting full GLOBOSA functional activity. Our results indicate that CsGLO1 and CsGLO2 paralogs might redundantly contribute to petal and stamen, providing the first insight into their role in tea plant flower development.

Antifungal mechanisms of α-terpineol and terpene-4-alcohol as the critical components of Melaleuca alternifolia oil in the inhibition of rot disease caused by Aspergillus ochraceus in postharvest grapes.

AIMS: As a natural antimicrobial agent, Melaleuca alternifolia oil (MAO) is generally recognized to be safe and effective in the inhibition of phytopathogenic fungi. Due to lack of comprehensive studies on MAO for controlling postharvest Aspergillus, we investigated the preservative mechanism of MAO and its components against Aspergillus ochraceus in postharvest grapes to evaluate their potential effectiveness as fruit preservatives. METHODS AND RESULTS: In our study, the compositions in MAO were analysed by gas chromatography-mass spectrometry. The inhibitory effects of MAO and its main constituents against A. ochraceus were compared by scanning electron microscopy and transmission electron microscopy observation, and metabolic analysis. Two components of MAO, α-terpineol and terpene-4-alcohol, showed higher antifungal effects than MAO, of which α-terpineol caused the worst leakage of cytoplasm and most serious hyphae distortions and spore disruptions. The downregulation of metabolic pathways of A. ochraceus was strongest with α-terpineol. The best inhibitory efficacy against A. ochraceus in grapes also occurred with α-terpineol. 3-Carene showed little inhibitory effect. CONCLUSIONS: These results demonstrate that not all components in MAO possess antimicrobial effects, and α-terpineol is the main contributor of MAO's A. ochraceus inhibition effect. SIGNIFICANCE AND IMPACT OF THE STUDY: α-Terpineol may be used as an alternative natural preservative for the postharvest storage of grapes and other fruits.

Antiviral activity of shikonin ester derivative PMM-034 against enterovirus 71 in vitro.

Human enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease (HFMD), particularly in infants and children below 4 years of age. Shikonin is a bioactive compound with anti-inflammatory, antiviral, and antibacterial activities derived from the roots of the Chinese medicinal herb Lithospermum erythrorhizon. This study aimed to examine the antiviral activity of PMM-034, a shikonin ester derivative, against EV71 in rhabdomyosarcoma (RD) cells. Cytotoxicity of PMM-034 on RD cells was determined using WST-1 assay. Dose- and time-dependent effects of PMM-034 on EV71 replication in RD cells were determined using plaque reduction assay. mRNA expression levels of EV71/VP1 and pro-inflammatory cytokines (IL-1ß, IL-6, IL-8, and TNF-α) were determined by real-time RT-PCR, and EV71/VP1 and phospho-p65 protein expressions were determined by western blot analysis. PMM-034 exhibited only weak cytotoxicity against RD cells. However, PMM-034 exhibited significant antiviral activity against EV71 in RD cells with 50% inhibitory concentration of 2.31 µg/mL. The VP1 mRNA and protein levels were significantly reduced in cells treated with PMM-034. Furthermore, relative mRNA expression levels of IL-1ß, IL-6, IL-8, and TNF-α significantly decreased in the cells treated with PMM-034, while the phospho-p65 protein expression was also significantly lower in the treated cells. These results indicated that PMM-034 suppressed the expressions of pro-inflammatory cytokines in RD cells, exhibiting antiviral activity against EV71, as evidenced by the reduced VP1 mRNA and protein levels in PMM-034-treated cells. Thus, PMM-034 is a promising candidate for further development as an EV71 inhibitor.

Antiviral activity of shikonin ester derivative PMM-034 against enterovirus 71 in vitro

Human enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease (HFMD), particularly in infants and children below 4 years of age. Shikonin is a bioactive compound with anti-inflammatory, antiviral, and antibacterial activities derived from the roots of the Chinese medicinal herb Lithospermum erythrorhizon. This study aimed to examine the antiviral activity of PMM-034, a shikonin ester derivative, against EV71 in rhabdomyosarcoma (RD) cells. Cytotoxicity of PMM-034 on RD cells was determined using WST-1 assay. Dose- and time-dependent effects of PMM-034 on EV71 replication in RD cells were determined using plaque reduction assay. mRNA expression levels of EV71/VP1 and pro-inflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) were determined by real-time RT-PCR, and EV71/VP1 and phospho-p65 protein expressions were determined by western blot analysis. PMM-034 exhibited only weak cytotoxicity against RD cells. However, PMM-034 exhibited significant antiviral activity against EV71 in RD cells with 50% inhibitory concentration of 2.31 μg/mL. The VP1 mRNA and protein levels were significantly reduced in cells treated with PMM-034. Furthermore, relative mRNA expression levels of IL-1β, IL-6, IL-8, and TNF-α significantly decreased in the cells treated with PMM-034, while the phospho-p65 protein expression was also significantly lower in the treated cells. These results indicated that PMM-034 suppressed the expressions of pro-inflammatory cytokines in RD cells, exhibiting antiviral activity against EV71, as evidenced by the reduced VP1 mRNA and protein levels in PMM-034-treated cells. Thus, PMM-034 is a promising candidate for further development as an EV71 inhibitor.

Diversity and antimicrobial activity of endophytic fungi isolated from Cephalotaxus hainanensis Li, a well-known medicinal plant in China.

UNLABELLED: About 1051 endophytic fungi were isolated from leaves, branches, barks and stems of Cephalotaxus hainanensis Li from four sites in Hainan, China. The fungi were identified as 21 genera by morphology and ITS sequences. One dominant species was Phomopsis quercella in Hainan Tropical Botanical Garden and Bawangling Nature Reserve, with relative frequency of 42·06 and 34·88% respectively. Another dominant species was Colletotrichum boninense in Wuzhishan and Jianfengling Nature Reserves, with relative frequency of 36·84 and 46·97% respectively. Among the selected 21 endophytic fungi, 17 strains (80·95%) had activity against at least one pathogenic bacteria, and 14 strains (66·67%) exhibited activity against at least one fungal pathogens. Neonectria macroconidialis showed strong inhibition against Staphylococcus aureus (inhibition zone being 20 mm), Bacillus subtilis (14 mm) and Streptococcus agalactiae (28 mm). Xylaria sp. showed strong inhibition against Escherichia coli (20 mm), Rhizoctonia solani (20 mm) and Sclerotinia sclerotiorum (17 mm). Verticillium bulbillosum showed great activity against Strep. agalactiae (32 mm) and Fusarium oxysporum (22 mm). These endophytic fungi showed potentials in medicine development. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic fungi from medicinal plants are an important source of novel and viable drugs. Cephalotaxus hainanensis Li is well known for leukaemia treatment and its endophytic fungi were isolated to investigate the diversity and antimicrobial activity. It was found that Ce. hainanensis Li had rich endophytic fungi, and some fungi showed strong antimicrobial activity against certain pathogens. These fungi can be used in medicine development.

Measurement of two low-temperature energy gaps in the electronic structure of antiferromagnetic USb2 using ultrafast optical spectroscopy.

Ultrafast optical spectroscopy is used to study the antiferromagnetic f-electron system USb(2). We observe the opening of two charge gaps at low temperatures (

Uranium and thorium leached from uranium mill tailing of Guangdong Province, China and its implication for radiological risk.

The paper focused on the leaching behaviour of uranium (U) and thorium (Th) from uranium mill tailing collected from the Uranium Mill Plant in Northern Guangdong Province, China. Distilled water (pH 6) and sulphuric acid solution (pH 4 and 3) were used as solvent for the leaching over 22 weeks. It was found that the cumulative leach fraction from the mill tailing was 0.1, 0.1 and 0.7 % for U release, and overall 0.01 % for Th release, using distilled water, sulphuric acid solution of pH 4 and pH 3 as leaching agents, respectively. The results indicate that (1) the release of U and Th in uranium mill tailing is a slow and long-term process; (2) surface dissolution is the main mechanism for the release of U and Th when sulphuric acid solution of pH 3 is employed as the leaching agent; (3) both U and Th are released by diffusion when using sulphuric acid solution of pH 4 as the leaching agent and (4) U is released by surface dissolution, while Th is released by diffusion when using distilled water as the leaching agent. The implication for radiological risk in the real environment was also discussed.

Baifuzi reduces transient ischemic brain damage through an interaction with the STREX domain of BKCa channels.

Stroke is a long-term disability and one of the leading causes of death. However, no successful therapeutic intervention is available for the majority of stroke patients. In this study, we explored a traditional Chinese medicine Baifuzi (Typhonium giganteum Engl.). We show, at first, that the ethanol extract of Baifuzi exerts neuroprotective effects against brain damage induced by transient global or focal cerebral ischemia in rats and mice. Second, the extract activated large-conductance Ca(2+)-activated K(+) channel (BK(Ca)) channels, and BK(Ca) channel blockade suppressed the neuroprotection of the extract, suggesting that the BK(Ca) is the molecular target of Baifuzi. Third, Baifuzi cerebroside (Baifuzi-CB), purified from its ethanol extract, activated BK(Ca) channels in a manner similar to that of the extract. Fourth, the stress axis hormone-regulated exon (STREX) domain of the BK(Ca) channel directly interacted with Baifuzi-CB, and its deletion suppressed channel activation by Baifuzi-CB. These results indicate that Baifuzi-CB activated the BK(Ca) channel through its direct interaction with the STREX domain of the channel and suggests that Baifuzi-CB merits exploration as a potential therapeutic agent for treating brain ischemia.

[Genetic transformation of hairy roots in Trichosanthes kirilowii Maxim. by Ti and Ri plasmids].

OBJECTIVE: To establish a hairy root culture system by double transformation for Trichosanthes kirilowii. METHOD: 1. Crown galls were induced by direct infection of sterile seedlings of T. kirilonii with Agrobacterium tumefaciens C58, and then the hairy roots were obtained from the regenerated plants by infection with A. rhzogenes 15834; 2. Transformation of Ti and Ri plasmids was inspected by high-pressure-paper electrophoresis; 3. The protein contents in the tissues of T. kirilowii were inspected by spectrophotometer and SDS-PAGE. RESULT: A hairy root culture system has been established successfully by double transformation with Ti and Ri plasmids in T. kirilowii. CONCLUSION: Compared with the ordinary hairy roots, the double transformed hairy roots grow faster but retain similar protein contents.

[Dynamics of growth and total tanshinones accumulation in crown gall cultures of salvia miltiorrhiza].

AIM: To determine the dynamics of growth and total tanshinones accumulation in crown gall cultures of Salvia miltiorrhiza in MS and 67-V liquid media. METHODS: Fresh, dry weight and total tanshinones yields in the cultures and in the medium were determined every 5 days in crown gall suspension cultures. RESULTS: In MS medium, the logarithmic growth phase of crown gall cultures in S. miltiorrhiza was from the 5th to 30th days, and the stationary growth phase was from the 30th to 35th days. From the 25th to 30th days, physiological activity of crown gall cultures was higher and their growth was better. However, in 67-V medium, the logarithmic growth phase of crown gall cultures was from the 10th to 25th days, and the stationary growth phase was from the 25th to 35th days. Total tanshinones were largely accumulated in the cultures and in the medium after 25 days. The total tanshinones yield (60 mg.L-1) was reached at the 35th day. CONCLUSION: Knowing the regularity of the growth and total tanshinones accumulation in crown gall cultures of S. miltiorrhiza will be helpful to take proper regulative measures in order to obtain the maximum total tanshinones yield.

Differentiation in a rat PC12 cell line induced by ostruthin and (-)-bornyl ferulate, constituents of a Chinese herbal medicine.

A search for neuritogenic compounds in Chinese herbs resulted in the isolation of two known substances, ostruthin and (-)-bornyl ferulate, from Notopterygium incisum (and/or N. forbesii). Both compounds induced comparable neurite-like structures in 20% of rat PC12 cells at 2 micrograms/ml, and showed cytotoxicity at concentrations higher than 3 micrograms/ml.

The immunostimulatory effect of bio-active peptide from pollen on murine and human lymphocytes.

The effects of bio-active peptide (BP) from buckwheat pollen and its synthetic analogues on murine and human lymphocytes and HL-60 cells were examined. Both natural and synthetic BP had the same effect in the lymphocyte blast transformation test. The proliferative response of HL-60 cells, co-cultured for 72 h with BP, was measured by the MTT method. BP had a stimulatory effect on HL-60 cell proliferation. The optical density (OD) value was 0.339+/-0.015, whereas, that of the control without BP was significantly lower (0.279+/-0.037; P < 0.05). The CD25 (IL-2Ralpha chain) positive cells of BP-stimulated human lymphocytes assessed by indirect immunofluorescence were 53.97+/-7.12% (1 microg/ml) and 42.53+/-4.4% (10 microg/ml), significantly higher than the control (11.77+/-1.88%; P < 0.001). The content of sIL-2R in the supernatant after culture with BP was determined by ELISA and was 1782.3+/-78.18 U/ml, lower than control 2381.2+/-99.9 U/ml. Taken together, our results showed that BP has an immunostimulatory effect on lymphocytes. The likely mode of BP action and the possibility of developing a novel immunomodulator are discussed.

[The plant regeneration of Salvia miltiorrhiza Bge. transformed by Agrobacterium].

The hairy roots and crown galls of Salvia miltiorrhiza were obtained by infecting plant with A. rhizogenes (strain 15834, LBA 9402) and A. tumefaciens (strain C58). The transformed plants were regenerated light and transplanted form cultural medium into soil successfully. The plants transformed by A. rhizogenes have characteristics of short stems and develop hairy roots, those and trans formed by A. tumefaciens grow vigorously featuring, longer stems and well developed roots. Both biomass production and tanshenone content are higher than the original plant.

Pharmacokinetic study on acetaminophen: interaction with a Chinese medicine.

To know the influences of a Chinese traditional medicine (KAKKONTO) on the metabolism of acetaminophen (APAP), we have carried out pharmacokinetic studies on APAP under KAKKONTO coadministration in humans and rats. In humans, the pharmacokinetic parameters were calculated from the blood APAP concentration-time curves of each volunteer. The parameters did not show any significant differences between the KAKKONTO-coadministration group (group K) and the APAP-administration group (group A). KAKKONTO, when given at two different doses, did not show any significant effects on blood APAP level. In rats, the blood APAP level was significantly higher than that of group A (p < 0.01) only in the 100 mg/kg of group K at 0.25 h after APAP administration. There were no other significant differences.

Selection of a high tanshinone-producing crown gall strain and production of tanshinone in the strain.

Crown galls were induced by direct infection of sterile seedlings of Salvia miltiorrhiza with Agrobacterium tumefaciens C58 and subcultured on a 67-V hormone-free medium by successively selecting red cell aggregates. A high tanshinone-producing crown gall strain C1 was obtained after 12 months in a subculture. It grows well and retains its high tanshinone-producing characteristic in liquid stationary cultures. It is obvious that the yeast extract and fermentation extract of Armillaria mellea as elicitors promote strain C1 to produce tanshinone. Tanshinone content of strain C1 cultures was over three times higher than that of the crude drugs. The results indicated that the crown gall tissue and elicitor technique could provide some new clues for the production of tanshinone under the condition of a liquid stationary culture.