Screening of an Armillaria gallica strain for Gastrodia elata cultivation / 中国中药杂志

This study aims to screen a strain from Armillaria for the cultivation of Gastrodia elata. Specifically, Armillaria strains were isolated from different producing areas of G. elata and identified. Based on the growth characteristics of the strains and the experiment on the cultivation of G. elata, an optimal A. gallica strain was screened out. The specific process is as follows. The fungus-gro-wing materials of G. elata were collected from four producing areas and the Armillaria strains were isolated(G,Y,S,H). The strains were then identified based on morphological observation and phylogeny analysis and the commonly used strains were determined. The sucrase genotypes of the strains were identified according to our previous research findings, and the growth characteristics of the strains, such as growth rate, diameter, dry weight, and polysaccharide content of the rhizomorphs, were measured. According to the biological characteristics and sucrase genotypes, two strains were selected for the cultivation of G. elata. The tuber yield and the content of gastrodin and p-hydroxybenzyl alcohol in the tuber of G. elata were measured to select the optimal strain. The results showed that the four strains were all A. gallica. The rhizomorphs of strains G and H of the same sucrase genotype had larger/higher length, growth rate, diameter, branch number, dry weight, and polysaccharide content than those of strains S and Y of the same sucrase genotype. The tuber yield and the total content of gastrodin and p-hydroxybenzyl alcohol in tuber of G. elata cultivated with strain H were 6.528 kg·m~(-2) and 0.566%, respectively, which were 4.58 and 1.30 folds those of G. elata cultivated with strain S. Strains H and S were screened out from four strains of A. gallica based on the growth characteristics and sucrase genotype. According to the tuber yield and content of total gastrodin and p-hydroxybenzyl alcohol in the tuber of G. elata, strain H was identified as the optimal one. The findings in this study are expected to lay a basis for cultivating G. elata with high yield and quality of tubers.

A Soybean Straw Cultivation Medium for Armillaria gallica and Optimal Composition / 中国实验方剂学杂志

Objective:To explore the feasibility of replacing wood （or wood chips） with crop residues for culturing <italic>Armillaria gallica</italic> targeting the problems of forest resource destruction and increased cultivation cost caused by the extensive use of wood in <italic>Gastrodia elata</italic> cultivation， so as to reduce the cultivation cost of <italic>G. elata</italic>， promote the effective use of crop residues， and protect forest resources. Method:The growth situation of <italic>A. gallica</italic> in different media was observed， followed by the measurement of its growth rate using streaking method and the determination of total polysaccharide content of <italic>A. gallica</italic> by phenol-concentrated sulfuric acid colorimetric method. In order to further optimize the soybean straw cultivation medium， we carried out a four-factor three-level L₉（3⁴） orthogonal assay on the ratio of main ingredients， sucrose content， inorganic salt content， and water content. Result:The comparison of growing states of <italic>A. gallica</italic> cultured in different media revealed that <italic>A. gallica</italic> in soybean straw medium began to grow since the fourth day of inoculation， and the mycelium grew well， with the growth rate being 0.352 cm·d^-1， which was 1.48 times that in birch wood medium. The total polysaccharide content of <italic>A. gallica</italic> cultured in soybean straw medium was the highest， which was 39.260 mg·g^-1， much higher than 17.028 mg·g^-1 of that cultured in birch wood medium. This demonstrated the obvious advantage of soybean straw medium， whose main ingredients were soybean straw and wheat bran at the ratio of 8：2， with the sucrose and inorganic salt content accounting for 1% and 0.5% of the main ingredients， respectively. When the water content reached 50%， the growth rate of <italic>A. gallica</italic> was maintained at 0.392 cm·d^-1. Conclusion:This study has provided a basis for utilizing soybean straw instead of wood （or wood chips） as cultivation medium for <italic>A. gallica</italic>， thus better reducing the waste of forest resources and protecting the natural environment in the cultivation of <italic>G. elata</italic>.

Effects of Exogenous Substances on Growth of Polyporus umbellatus Mycelium and Its Polysaccharide Content / 中国实验方剂学杂志

Objective:To explore the effects of diverse exogenous substances at different concentrations on the growth of<italic> Polyporus umbellatus</italic> mycelium and polysaccharide content and screen out the optimal growth condition for <italic>P. umbellatus</italic> mycelium， so as to provide a reference for its large-scale artificial cultivation. Method:<italic>P. umbellatus</italic> mycelium was cultured in media containing different exogenous substances using the method for fungal culturing in plate. The growth rate of the mycelium was judged by the colony diameter and the polysaccharide content was determined by the phenol-sulfuric acid method. Result:The high-dose cyclic adenosine monophosphate， 6-benzyl aminopurine （6-BA）， gibberellic acid （GA）， 2，4-dichlorophenoxyacetic acid （2，4-D）， vitamin （V） B₁， VB₃， VB₆， VB₉， and VB₁₂ all promoted the growth of <italic>P. umbellatus</italic> mycelium and elevated polysaccharides content. By contrast， indole acetic acid （IAA）， VC， and VB₂ inhibited its growth， with the most obvious inhibition detected in the high-dose VC group. IAA and VB₂ both reduced the polysaccharide content， whereas the high-dose VC significantly increased the polysaccharide content. Cyclic adenosine monophosphate， 6-BA， GA， 2，4-D， VB₁， VB₃， VB₆， VB₉， and VB₁₂ at the concentrations of 2 mmol·L^-1， 6 mg·L^-1， 15 mg·L^-1， 2 mg·L^-1， 4 mg·L^-1， 2 mg·L^-1， 4 mg·L^-1， 6 mg·L^-1， and 10 mg·L^-1， respectively， contributed to the growth of <italic>P. umbellatus</italic> mycelium<italic> </italic>and polysaccharide accumulation. Conclusion:The growth of <italic>P. umbellatus </italic>mycelium and polysaccharide accumulation can be regulated by adding exogenous substances to the culture medium.

Identification of Gastrodia elata and its hybrid by polymerase chain reaction / 中国中药杂志

Gastrodia elata is a kind of traditional Chinese medicinal materials and has good medicinal value. G. elata is divided into five varieties, which includes G. elata f. elata(proto variant), G. elata f. glauca, G. elata f. viridis, G. elata f. flavid and G. elata f. alba. Among them, G. elata f. elata and G. elata f. glauca have excellent characteristics and higher contents of gastrodin and polysaccharides. The hybrid of G. elata f. elata and G. elata f. glauca is present in markets, but the characteristics between hybrid and parent are not obvious and distinguished quickly and accurately. The aim of this study is to establish a PCR specific PCR identification method, which can identify G. elata f. elata, G. elata f. glauca and their hybrid. Based on the re-sequencing results of G. elata, we screened for the single nucleotide polymorphism(SNP) variation sites, and designed two pairs of specific primers(W291-F/W291-R and H255-F/H255-R). We further collected G. elata f. elata, G. elata f. glauca and their hybrid samples from different regions, established and optimized PCR method, and investigated and verified their tolerance and applicability. The results showed that when the annealing temperature was 48 ℃ and the number of cycles was 33, 255 bp specific band were obtained from G. elata f. glauca and hybrid by using specific primers W291-F/W291-R. When the annealing temperature was 51 ℃ and the number of cycles was 33, 291 bp specific band were obtained from G. elata f. elata and hybrid by using specific primers H255-F/H255-R. Our method could be used as a promising method to identify G. elata f. elata, G. elata f. glauca and their hybrid.

Survey on Woods Species and Its Relation with Yield and Quality of Gastrodia elata / 中国实验方剂学杂志

Objective:As a source of energy for Armillaria mellea and Gastrodia elata，the woods species as fungus material of G.elata are diverse and play an important role in the development of G.elata industry. In order to explore the impact of different woods species on the quality of G. elata，the plant origins and lignocellulose content of the woods，the yield and quality of G. elata per unit area were systemically analyzed through literature research and investigation on production bases. Method:G. elata and its cultivated woods were collected from four main producing areas （Guizhou, Yunnan, Hubei, and Shaanxi），and the the plant origins of the woods were identified by DNA fragments. The content of lignocellulose in the woods was determined by high-performance liquid chromatography and loss-on-ignition method. The content of polysaccharides of G. elata from these 4 areas was determined by Phenol-sulfuric acid method. The yields and polysaccharide content of G. elata cultivated with different woods species were compared and their correlation with the woods was analyzed. Result:The woods as fungus material of G. elata were diverse in species, and betulaceae was the most widely used species in cultivation of G. elata. There were differences in the composition ratio of lignocellulose in the woods. Nyssaceae had the highest cellulose content，Moraceae had the highest hemicellulose content and Rosaceae had the highest lignin content. Different woods species had certain effects on the yield and polysaccharide content of G. elata. The maximum yield of G. elata was 1 285.51 g and the lowest yield was 379.30 g. The average mass fraction of polysaccharide content was 241.1 mg·g-1，with a range of 87.95-411.2 mg·g-1. The yield and polysaccharide content of G. elata were highly positively correlated with the cellulose content of the woods, and highly negatively correlated with the lignin content. Conclusion:Different woods have a significant impact on the yield and quality of G. elata. Choosing the appropriate woods species will be beneficial to the absorption of nutrients for A. mellea and the yield increase of G. elata. This study can provide a scientific basis for the selection of woods species during the cultivation of G. elata.

Observation on effects of hepatitis B vaccine immunization for 12 years in children in Beijing / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate the long-term protective effects of hepatitis B vaccine after immunizing to the children for 12 years in Beijing.</p><p><b>METHODS</b>The multiple stratified cluster sampling was used in this epidemiological survey. The sampling children's blood serum HBsAg, anti-HBs and anti-HBc were checked and measured by the solid phase radioimmunoassay (SPRIA). The serological level of these index and the causes of the children with HBsAg positive were analyzed.</p><p><b>RESULTS</b>There were 2,419 cases 3-12 years-old children immunized with the hepatitis B vaccine in infant period were surveyed and the total HBsAg positive rate was 0.52%. The vaccine protective rate was 88.45% (95% CI: 65.67%-97.89%). The total anti-HBc positive rate was 2.21%, being no statistical significance among the age groups. The average anti-HBs positive rate of 3-6 years-old children immunized with gene recombining vaccine was 38.79% and descending greatly following the age's dropping. The geometric means of anti-HBs serological titer (GMT) was 52.83 mIU/ml, showing no statistical significance among the age groups. The average anti-HBs positive rate of 6-12 years-old children immunized with the blood rooting vaccine was 50.79%. The geometric means of anti-HBs serological titer (GMT) was 61.51 mIU/ml. There were no statistical significances among the age groups. Among the HBsAg positive children, more than 50% of the children's mothers were HBsAg positive also.</p><p><b>CONCLUSIONS</b>The protective effects given by immunization were significant after the hepatitis B vaccine vaccination for 12 years in Beijing. The booster immunization was not necessary, because the HBsAg positive rate didn't ascend obviously as the immunization time prolonging. As the anti-HBs positive rate of children who were immunized by the gene recombining vaccine might be descending following the age's dropping greatly, we should strengthen the serological surveillance of hepatitis B. The main cause that the children became the HBsAg carrier should be a vertical transmission.</p>

Study on the afficacy of hepatitis B immunization among youngsters in Beijing / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluating the epidemiological effect after hepatitis B immunization among youngsters in Beijing.</p><p><b>METHODS</b>A multistage sampling method was used for data collection from immunization cards, reports on field epidemiological survey on hepatitis B virus (HBV) immunization of youngsters and the analysis of infectious diseases. HBsAg, anti-HBs and anti-HBc levels were detected by solid phase radioimmunoassays (SPRIA).</p><p><b>RESULTS</b>The average incidence rates of hepatitis B in 10 - 19 years-olds prior to HBV immunization was 12.11-16.89/100 000, while declined to 1.78-10.95/100 000 (chi(2) = 10.71 - 60.45, P < 0.01). HBsAg carrying rate of the youngsters decreased from 6.34% to 1.37% (80.00%) after vaccination (chi(2) = 14.16, P < 0.01).</p><p><b>CONCLUSION</b>Hepatitis B vaccination among youngsters is an effective measure for the prevention and control of hepatitis B virus infection.</p>

Epidemiological effect of hepatitis B immunization among newborn babies in Beijing / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the epidemiological effect of hepatitis B immunization among newborn babies in Beijing.</p><p><b>METHODS</b>A multistage sampling method was used for the collection of immunization cards, field epidemiological survey on hepatitis B virus (HBV) immunization of children, analysis of infectious disease reports. HBsAg, anti-HBs and anti-HBc levels were detected by solid phase radioimmunoassays (SPRIA).</p><p><b>RESULTS</b>The incidence of hepatitis B in children of 0 to 14 years at the beginning of HBV immunization was 18.59 to 20.52/100,000, and declined to 0.39 to 2.38/100,000 in 2000 (chi2 = 58.26, P < 0.01). The HBsAg carrying rate of the children decreased from 2.82% to 0.60%, about 80.00% after vaccination (chi2 = 10.75, P < 0.01).</p><p><b>CONCLUSION</b>Hepatitis B vaccination of newborn babies is an effective measure for prevention and control of hepatitis B virus infection.</p>