[Viability of 7 kinds of medicinal plant seeds stored in medium-term gene bank of the National Medicinal Plant Gene Bank].

In order to evaluate seed viability of Platycodon grandiflorum, Schizonepeta tenuifolia, Andrographis paniculat, Codonopsis pilosula, Scutellaria baicalensis, Leonurus japonicus, Rabdosia rubescens, stored in the medium-term gene bank of the National Medicinal Plant Gene Bank for 4 years, we tested seed germination rate of 7 species of medicinal plant and analyzed the change of significance of levels of the germination rate in pre and post store. Seed germination rates of 7 species of medicinal plants were all decreased after 4 years, and the decrease of S. tenuifolia and S. baicalensis germination rates were much smaller than other species. The higher initial germination rate of P. grandiflorum, C. pilosula, R. rubescens seed has the smaller decline of germination rate, but the data of A. paniculata showed the opposite trend. The rate decline of the germination of S. tenuifolia and S. baicalensis was roughly the same in different germination rate interval. The results showed that low temperature storage could effectively prolong the seed longevity, and maintain the seed vigor. Moreover, it is necessary to study on the storage characteristics of the main medicinal plant seeds, and establish the monitoring plan and regeneration standard.

[Variation and correlation analysis on phenotypic traits of Scutellaria baicalensis from different strains].

To research the differences and correlation between Scutellaria baicalensis about phenotypic traits of different strains, 10 aboveground traits and 6 root traits of S. baicalensis in two-year-transplanted plants from 14 different strains were compared respectively, and the SPSS 17.0 statistical software was used for data analysis. It showed that phenotypic traits variation of different S. baicalensis strains was rich and the F value ranged from 3.169 to 71.58. The difference was significant between each other and germplasm 15 performs the most outstanding characters. Correlation analysis showed that there existed a significant correlation between the characters except for lateral root number, root diameter and length. The correlation coefficient between the fresh weight of root and the reed head diameter was up to 0.877. Principal component analysis showed that the average of overall yield per plant and root diameter could be used as the comprehensive reference index for germplasm evaluation. The differences and correlations in phenotypic traits of different S. baicalensis strains, provide theoretical basis for distinguishing germplasm and breeding good varieties of S. baicalensis.

[Establishment of a cell suspension culture system of endangered Aquilaria sinensis (Lour.) Gilg].

Aquilaria sinensis callus induced by stem tips were used to establish the suspension cell system. The results showed that the most suitable medium for callus induction and subculture is MS + 2.0 mg x L(-1) NAA + 1.0 mg x L(-1) 6-BA. After 12 times of subculture, the energetic and loose callus, which were appropriate for cell suspension culture, were cultured and shook in liquid medium MS + 2.0 mg x L(-1) NAA + 1.0 mg x L(-1) 6-BA + 500.0 mg x L(-1) casein hydrolysate (CH) to establish the suspension cell system. The growth curve of suspension cells showed a "S" type. At the beginning of the culture, cell density increased slowly; during 4 to 6 days, suspension cells reached logarithmic growth period; during 7 to 12 days, suspension cells were in the platform period; but after 12 days, cell density and activity went down obviously. Agarwood sesquiterpenes were not detected in the suspension cells during the growth period, however, they could be detected in MeJA treated suspension cells. In this study, a stable and active growing suspension cell system was established, which was a proper system to study the mechanism of agarwood sesquiterpene formation, and additionally provided a potential way to generate agarwood sesquiterpenes through application of cell culture.

[Isolation and identification of pathogen of seedling blight of Platycodon grandiflorum].

OBJECTIVE: To isolate and identify pathogen of the seedling blight occurred in Platycodon grandiflorum. METHOD: The morphological observation, rDNA ITS sequence analysis, and Koch's postulates were used to identify the isolates of the causal agent. RESULT: The isolates of the causal agent was Rhizoctonia solani. CONCLUSION: The result confirmed that R. solani is the pathogen of seedling blight of P. grandiflorum.

[Studies on high efficient plant regeneration system from anther callus of Bupleurum chinense].

The callus of Bupleurum chinense with anthers at the stage of uninucleate was induced. After several subcultures, anther calli of B. chinense were cultured at 20 MS culture mediums with different plant hormones to differentiate into plantlets. Differentiation of callus was detected after 21 and 49 days to select the most effective medium. There were 19 culture mediums in which anther callus could differentiate into plantlets with differentiation rate range from 3% to 60% , and most less than 20%. MS + KT 0.5 mg x L(-1) + sucrose 30 g c L(-1) + phytagel 5 g x L(-1) was the best differentiation medium with the differentiation rate of 60%, followed by MS + ZT 1.0 mg x L(-1) + sucrose 30 g x L(-1) + phytagel 5 g x L(-1) with the differentiation rate of 58%. Then plantlets were transferred to rooting medium to obtain whole plant. All plantlets could root in the rooting medium of MS + sucrose 30 g x L(-1) + phytagel 5 g x L(-1) and 1/2 MS + NAA 0.5 mg x L(-1) + sucrose 30 g x L(-1) + phytagel of 5 g L(-1) with the rooting rate of 100%. As a result, the high efficient and stable plant regeneration system was established from anther callus of B. chinense.

Cardioprotective effect of a hemoglobin-based oxygen carrier on cold ischemia/reperfusion injury.

OBJECTIVES: The etiology of myocardial ischemia/reperfusion (I/R) injury is multifactorial, but activation of the innate immune system and the resulting inflammatory response are important components of I/R injury. The aim of this study was to investigate the protective effect of a hemoglobin-based oxygen carrier (HBOC) on cold I/R heart and to explore the underlying mechanisms. METHODS: Isolated Sprague-Dawley rat hearts were perfused in the Langendorff mode. After 30 min of basal perfusion, rat hearts were arrested with histidine-tryptophan-ketoglutarate solution (HTKs) with or without an HBOC and hypothermically stored (4°C) for 9 or 14 h, followed by 2 h of reperfusion. RESULTS: Compared with HTKs alone, the HBOC in HTKs greatly improved heart contraction and decreased infarct size, necrosis and apoptosis, which was related to the reduced expression of Toll-like receptor 2 (TLR 2), TLR 4, TNF-α, IL-1ß and nuclear factor-κB (NF-κB) activation. CONCLUSIONS: Our results demonstrated that the HBOC protected isolated rat heart from cold I/R injury and this protection was associated with attenuation of the expression of the TLR 2 and TLR 4/NF-κB signaling pathway, which may down-regulate the inflammatory response.

Polymerised placenta haemoglobin attenuates cold ischaemia/reperfusion injury in isolated rat heart.

Ischaemia/reperfusion (I/R) injury is harmful to the cardiovascular system and is responsible for the inflammatory response, which, in turn, aggravates cardiac dysfunction. This study was designed to investigate the protective effect and potential mechanism of a haemoglobin-based oxygen carrier on cold I/R-injured hearts. Isolated Sprague-Dawley rat hearts were perfused in Langendorff mode. After a 30-min basal perfusion, rat hearts were arrested and hypothermically stored at 4°C for 12h followed by a 2-h reperfusion. Compared with histidine-tryptophan-ketoglutarate solution (HTKs), polymerised placenta haemoglobin (PolyPHb) in HTKs greatly improved heart contraction and decreased infarction size, necrosis, and apoptosis, which was related to reduced expression of TLR2, TLR4, TNF-α, and IL-1ß, and NF-κB activation. Our results demonstrate the cardioprotective effect of PolyPHb on cold I/R-injured hearts and revealed that this protection was mediated in large part by attenuation of TLR2 and -4/NF-κB signalling pathway and could possibly down-regulate the inflammatory response.

[Comparison of the contents of mineral elements in the roots of four germplasms Bupleurum L. planted in different soils].

Contents of 21 elements, including of K, Ca, Mg, Na, Fe, Mn, Cu, Zn, Ni, Ti, Cr, Mo, V, Zr, Co, Se, Sn, Cs, Cd, Hg, and Pb, in the roots of four germplasms Bupleurum L. planted in the soils from seven regions, were analyzed by ICP-MS/ICP-AES. The results showed that the contents of K, Ca, Mg and Na are mainly ascribed to the effect of germplasms, and the contents of Fe, Cu, Ni, Ti, Cr, V, Zr, Se, Sn, Cs, Cd, Hg and Pb are mainly ascribed to the effect of soils considering the integrated effects of germplasms and soils. Generally, element contents in the root of G-GS were higher and that in the root of G-ZC were lower in the four germplasms of Bupleurum L. Element contents in the root of Bupleurum L. planted in S-HR were higher, while that planted in SYQS were lower as a whole among these soils from seven regions. The difference in the element contents has an important effect on the research into the contents of officinal ingredients and their curative effect in the roots of four original Bupleurum L. germplasms planted in the soils from seven regions. And this study also provides an important scientific foundation for the choices of germplasms and soils in the cultivation of high-yield and high-quality Bupleurum L.

[Heterosis analysis of the main agronomic characters of Platycodon grandiflorum].

OBJECTIVE: Breeding the good varieties characters of Platycodon grandiflorum. METHODS: 20 combination, derived from 2 x 10 incomplete diallel cross with 2 male sterile lines and 10 four-generation inbred lines, analysis the 13 agronomic characters. RESULTS: The agronomic characters of F1 has obvious heterosis. Dry plant weight and fresh root weight have the largest heterosis. Their middle parent heterosis (MPH), high parent heterosis (HPH) and heterosis over CK (CH) were 96.07%, 62.62%, 116.21%, 108.99%, and 111.98%, 113.47%, respectively. CONCLUSION: The F1 hybrids derived from the crossing of male sterile lines and in-bred lines have obvious heterosis. It applied the hybrid heterosis breeding is an effective way for the new variety of Platycodon grandiflorum.

A novel dynamic heterogeneous phase polymerization reaction for poly-hemoglobin with narrow molecular weight distribution.

A dynamic heterogeneous phase polymerization reaction is found to be efficient for controllable cross-link of hemoglobin with glutaraldehyde. The selective absorption of the immobile phase and asymmetry of protein concentration leads to narrowness of the molecular weight distribution and lowness of the average molecular weight. Using this method, 53% of hemoglobin obtained is intermolecular cross-linked with 12 molecular equivalents of glutaraldehyde. The majority of poly-hemoglobins is in the range of 128 kD to 258 kD.

DBBF modified human placenta hemoglobin through the alpha chains.

Hemoglobin-based blood substitute is widely studied. As the starting materials, hemoglobin (Hb) is mainly supplied from outdated human blood and animal sources. But there are many disadvantages. As the alternative source, human placenta hemoglobin (PHb) has its own advantages. We take the lead in being occupied in the research of blood substitutes based on the placenta blood. Purified PHb readily undergoes dissociation to form two dimers that are easily filtered by the kidneys and have high 02 affinity (low P50), failing to transport 02 to tissues. As a first step, to enhance PHb stability and P50, we first adopted DBBF to modify deoxyPHb leading to DBBF-alpha PHb. According to detection, we knew that the PHb stability was improved, 02 affinity of the PHb solution was reduced, DBBF-alpha PHb had the appropriate capacity of oxygen-carrying and oxygen-unloading, and DBBF-alpha PHb kept the biological activity well. On the whole DBBF-alpha PHb could be considered as a first step toward the synthesis of a potential blood substitute.

[Observation of pollen tube germination of Platycodon grandiflorum by fluorescence microscopy].

OBJECTIVE: To determine the self-compatibility of Platycodon grandiflorum and the location of microspore germination on stigma. METHOD: The microspore germination of self-pollination, self-plant-pollination and cross-pollination and the pollination microspore germination in and outside of stigma were observed with fluorescence microscopy. RESULT: Most pollens of self-pollination, self-plant-pollination and cross-pollination can germinate on the stigma, and after 24 hours, pollen tube entered into the ovary successfully. Pollinated on the outer-surface of stigma, microspores could not germinate, but on the inner-surface of the stigma when it dehisced most microspores can germinate. CONCLUSION: The compatibility of self-plant-pollination of Platycodon grandiflorum is high. The microspore germination loci is on the inner-surface of the dehisced stigma.

[Study on the stigma/pollen vigor and self-compatibility of Platycodon grandiflorum].

OBJECTIVE: To provide the basal data for the breeding and cultivation of Platycodon grandiflorum. METHOD: The field investigation and pollination by bagging were carried out. TTC(2,3,5-triphenyl tetrazolium chloride) solution was used to test the pollen vigor. RESULT: The stigma life-span of P. grandiflorum was about 9 days, however the optimal time for pollination is 4-6 days after the petals opening, with the stigma was splitting lightly or significantly. When the petals opened, the anther began scattering pollen, and finished in the same day. The pollen vigor was about 81.4% at the beginning, and decreases to 27.6% three days later. The pollen vigor still remains 64.4% three days later, when the flower was kept in the desiccant. The natural fructification rate of self-flower-pollination was 4.8%. The fructification rate and compatible index was about 62.7% and 54.6, respectively, when the self-plant-pollination performed by hand. They decreased to 12.8% and 6.5 when the pollination was implemented during the flowering period. CONCLUSION: Low fructification percentage of self-flower-pollination attributes to the difference of maturing period of pistil and stamen, as well as the short pollen life-span of P. grandiflorum. The compatibility of self-plant-pollination is high during flowering period when pollination performed by hand. The life-span of the pollen can be prolonged significantly when keeping in the dry environment.

A method for purification and viral inactivation of human placenta hemoglobin.

For pilot-scale manufacturing of hemoglobin-based oxygen carrying drugs, we should get highly pure and viral inactivated hemoglobin (Hb) at high recovery. In our method, placenta hemoglobin (PHb) solutions were purified by heating in the presence of reducing agent and deoxygenating conditions so that heat-sensitive proteins were selectively precipitated and virus was inactivated. The optimum preparative condition resulted in highly purified PHb solution (>99% pure) with approximate 90% recovery and less than 2% of MetHb content, maintained oxygen carrying capacity, residual phospholipids less than 1 ppm, free of endotoxin, bacteria, type A&B antigens and virus. Finally, we compared the efficacy of blood exchange on rat with poly-PHb and poly-Hb from adult blood. The results showed no significant difference between two products. Therefore, the placenta Hb obtained from this method could be supplied as materials for oxygen carrying drugs.