How to improve CHMs quality: Enlighten from CHMs ecological cultivation.

Chinese herbal medicines (CHMs) are one of the important bioresources of medicine, which works by unlocking nature's ability to prevent diseases and recover from illnesses. Recently, it has ascended to the world stage and become a global icon. Nowadays, a considerable of researches have focused on the quality evaluation of CHMs. However, it is difficult to meet the reasonable needs of human beings for safe drug use to evaluate the quality of a huge number of inferior goods for the CHMs contaminated by pesticides and heavy metals. Hence to explore an eligible medicinal plant cultivation pattern, which can provide high quality CHMs sustainably, is most promising. This review analyzed the situation and characteristics of medicinal plant resources in different periods, including wild-harvested and cultivated resources during different stages, putting forward that ecological cultivation must be the way to develop medicinal plant cultivation and to obtain high quality CHMs.

Detection of Cistanches Herba (Rou Cong Rong) Medicinal Products Using Species-Specific Nucleotide Signatures.

Cistanches Herba is a medicinal plant that has tonification properties and is commonly used in Asia. Owing to the imbalance between supply and demand, adulterants are frequently added for profit. However, there is no regulatory oversight because quality control tools are not sufficient for identifying heavily processed products. Thus, a novel molecular tool based on nucleotide signatures and species-specific primers was developed. The ITS2 regions from 251 Cistanches Herba and adulterant samples were sequenced. On the basis of SNP sites, four nucleotide signatures within 30~37 bp and six species-specific primers were developed, and they were validated by artificial experimental mixtures consisting of six different species and different ratios. This method was also applied to detect 66 Cistanches Herba products on the market, including extracts and Chinese patent medicines. The results demonstrated the utility of nucleotide signatures in identifying adulterants in mixtures. The market study revealed 36.4% adulteration: 19.7% involved adulteration with Cynomorium songaricum or Cistanche sinensis, and 16.7% involved substitution with Cy. songaricum, Ci. sinensis, or Boschniakia rossica. The results also revealed that Cy. songaricum was the most common adulterant in the market. Thus, we recommend the use of species-specific nucleotide signatures for regulating adulteration and verifying the quality assurance of medicinal product supply chains, especially for processed products whose DNA is degraded.

Combined Modification of Dual-Phase Li4Ti5O12-TiO2 by Lithium Zirconates to Optimize Rate Capabilities and Cyclability.

The low electrical conductivity and ordinary lithium-ion transfer capability of Li4Ti5O12 restrict its application to some degree. In this work, dual-phase Li4Ti5O12-TiO2 (LTOT) was modified by composite zirconates of Li2ZrO3, Li6Zr2O7 (LZO) to boost the rate capabilities and cyclability. When the homogeneous mixture of LiNO3, Zr(NO3)4·5H2O and LTOT was roasted at 700 °C for 5 h, the obtained composite achieved a superior reversible capacity of 183.2 mAh g-1 to the pure Li4Ti5O12 after cycling at 100 mA g-1 for 100 times due to the existence of a scrap of TiO2. Meanwhile, when the composite was cycled by consecutively doubling the current density between 100 and 1600 mA g-1, the corresponding reversible capacities are 183.2, 179.1, 176.5, 173.3, and 169.3 mAh g-1, signifying the prominent rate capabilities. Even undergoing 1400 charge/discharge cycles at 500 mA g-1, a reversible capacity of 144.7 mAh g-1 was still attained, denoting splendid cyclability. From a series of comparative experiments and systematic characterizations, the formation of LZO meliorated both the Li+ migration kinetics and electrical conductivity on account of the concomitant superficial Zr4+ doping, responsible for the comprehensive elevation of the electrochemical performance.

Rapid Authentication of Ginkgo biloba Herbal Products Using the Recombinase Polymerase Amplification Assay.

Species adulteration in herbal products (HPs) exposes consumers to health risks. Chemical and morphological methods have their own deficiencies when dealing with the detection of species containing the same active compounds in HPs. In this study, we developed a rapid identification method using the recombinase polymerase amplification (RPA) assay to detect two species, Ginkgo biloba and Sophora japonica (as adulteration), in Ginkgo biloba HPs. Among 36 Ginkgo biloba HP samples, 34 were found to have Ginkgo biloba sequences, and 9 were found to have Sophora japonica sequences. During the authentication process, the RPA-LFS assay showed a higher specificity, sensitivity and efficiency than PCR-based methods. We initially applied the RPA-LSF technique to detect plant species in HPs, demonstrating that this assay can be developed into an efficient tool for the rapid on-site authentication of plant species in Ginkgo biloba HPs.

Detection of Ophiocordyceps sinensis and Its Common Adulterates Using Species-Specific Primers.

Ophiocordyceps sinensis is a fungus that infects Hepialidae caterpillars, mummifying the larvae and producing characteristic fruiting bodies (stromata) that are processed into one of the most valued traditional Chinese medicines (TCM). The product commands a very high price due to a high demand but a very limited supply. Adulteration with other fungi is a common problem and there is a need to test preparation for the presence of the correct fungus. In the current study, a PCR-based approach for the identification of O. sinensis based on a segment of the internal transcribed spacer (ITS) region was developed. The segments is 146-bp in size and is likely to be amplified even in materials where processing led to DNA fragmentation. Primer development was based on the alignment of sequence data generated from a total of 89 samples of O. sinensis and potential adulterants as well as sequences date from 41 Ophiocordyceps species and 26 Cordyceps species available in GenBank. Tests with primer pair, DCF4/DCR4, demonstrated generation of an amplicon from DNA extracted from O. sinensis stromata, but not from extracts derived from adulterants. Species-specific primer pairs were also developed and tested for detection of the common adulterants, Cordyceps gunnii, Cordyceps cicadae, Cordyceps militaris, Cordyceps liangshanensis and Ophiocordyceps nutans. The collection of primers developed in the present study will be useful for the authentication of preparation claiming to only contain O. sinensis and for the detection of fungi used as adulterants in these preparations.

Analysis of the seismic performance of isolated buildings according to life-cycle cost.

This paper proposes an indicator of seismic performance based on life-cycle cost of a building. It is expressed as a ratio of lifetime damage loss to life-cycle cost and determines the seismic performance of isolated buildings. Major factors are considered, including uncertainty in hazard demand and structural capacity, initial costs, and expected loss during earthquakes. Thus, a high indicator value indicates poor building seismic performance. Moreover, random vibration analysis is conducted to measure structural reliability and evaluate the expected loss and life-cycle cost of isolated buildings. The expected loss of an actual, seven-story isolated hospital building is only 37% of that of a fixed-base building. Furthermore, the indicator of the structural seismic performance of the isolated building is much lower in value than that of the structural seismic performance of the fixed-base building. Therefore, isolated buildings are safer and less risky than fixed-base buildings. The indicator based on life-cycle cost assists owners and engineers in making investment decisions in consideration of structural design, construction, and expected loss. It also helps optimize the balance between building reliability and building investment.

[Study on traceability system of genuine medicinal materials].

Genuine medicinal materials with special characteristics of Traditional Chinese Medicine (TCM), is recognized as high quality medicine. Both ancient records and modern research considered that the origin is an important reason for the formation of genuine medicinal materials. However, blindly transplanting of genuine medicinal materials has led to the quality decline and counterfeit medicines appeared in production or sale progress, which may increase the risk of accidents in TCM. Frequent accidents emerged in Chinese herbal affects its export. What's more, it is a great threat to the medication safety in TCM clinical. There is an urgent need to implement traceability systems of TCM, which could provide convenient information record and traceability of TCM circulation. This paper reviews a variety of technical methods for genuine medicinal materials traceability, and proposed the establishment of genuine medicinal materials traceability system based on two-dimensional code and network database.

[RNA-Seq and genuine traditional Chinese medicine].

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.

[Identification of Salvia shandongensis new species based on sequences of the plastid psbA-trnH intergenic region].

To identify Salvia shandongensis and its relatives at molecular level, the psbA-trnH intergenic region of three species including Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba were amplified and sequenced. Sequences were assembled with CodonCode Aligner. The K2P genetic distances between Salvia shandongensis and its relatives were calculated and UPGMA tree was performed by MEGA5.0. The results indicated that the lengths of psbA-trnH regions of Salvia shandongensis were about 391 bp, while the lengths of psbA-trnH regions of Salvia miltiorrhiza and S. miltiorrhiza f. alba were about 386 bp. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Salvia shandongensis and its relatives. The intra-specific genetic distances of Salvia shandongensis were 0, while the intra-specific genetic distances of Salvia miltiorrhiza and S. miltiorrhiza f. alba were 0.002 and 0.001 respectively. Additionally, the genetic distance of Salvia shandongensis and Salvia miltiorrhiza ranged from 0.034 to 0.04, and the genetic distance of Salvia shandongensis and S. miltiorrhiza f. alba ranged from 0.005 to 0.008, the intra-specific genetic distances of Salvia shandongensis were much smaller than that of Salvia miltiorrhiza and S. miltiorrhiza f. alba; clustering results showed that there were obvious differences between Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba, which was consistent with morphological characteristics. This study not only firstly provides the scientific basis for establishing the taxonomy position in molecular level and revealing their genetic relationships of S. shandongensis, S. miltiorrhiza and S. miltiorrhiza f. alba; but also provides DNA molecular identification scientific basis for the development of new medicinal plant resources of Salvia shandongensis. Our results suggest that the psbA-trnH intergenic spacer region can be used as a barcoding to identify Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba.

Molecular identification of Corni Fructus and its adulterants by ITS/ITS2 sequences.

UNLABELLED: The DNA barcoding method was used to accurately and rapidly identify Corni Fructus and its adulterants. METHODS: Genomic DNA extracted from Corni Fructus and its adulterants were used as templates. The ITS (internal trascribed spacer) regions were amplified using polymerase chain reaction. Sequence assembly was performed using CodonCode Aligner V 3.5.4. Genetic distances were computed using MEGA V 5.0. Species identification was conducted using neighbor-joining (NJ) trees. RESULTS: The ITS sequence length of Corni Fructus was 659 bp. The average intra-specific genetic distance of Corni Fructus was 0.005, markedly lower than the inter-specific genetic distance between Corni Fructus and its adulterants (0.357). The ITS2 sequence length of Corni Fructus was 250 bp. No variation was found among the different samples. The interspecific genetic distance of ITS2 between Corni Fructus and its adulterants was 0.571. NJ trees and BLAST results indicated that Corni Fructus and its adulterants can be easily differentiated with monophyly. CONCLUSION: ITS/ITS2 regions can accurately and efficiently distinguish Corni Fructus and its adulterants. In addition, the results not only established the foundation for the clinical safety in the utilization of Corni Fructus, but also provided reference for molecular identification of other Chinese herbal medicine and Chinese herbal pieces.

[Principles for molecular identification of traditional Chinese materia medica using DNA barcoding].

Since the research of molecular identification of Chinese Materia Medica (CMM) using DNA barcode is rapidly developing and popularizing, the principle of this method is approved to be listed in the Supplement of the Pharmacopoeia of the People's Republic of China. Based on the study on comprehensive samples, the DNA barcoding systems have been established to identify CMM, i.e. ITS2 as a core barcode and psbA-trnH as a complementary locus for identification of planta medica, and COI as a core barcode and ITS2 as a complementary locus for identification of animal medica. This article introduced the principle of molecular identification of CMM using DNA barcoding and its drafting instructions. Furthermore, its application perspective was discussed.

[Research progress on diagnostic methods for Babesia microti infection].

Human babesiosis, a malaria-like zoonosis transmitted by the tick, is mainly distributed in Europe, USA and some Asian countries. There are various kinds of diagnostic methods for babesiosis caused by Babesia microti, but many of them are still in the preliminary stage. This article reviews the main diagnostic techniques and the existing problems.

[Stability and accuracy of the identification of Notopterygii Rhizoma et Radix using the ITS/ITS2 barcodes].

In this study, Notopterygii Rhizoma et Radix was used to verify the stability and accuracy of DNA barcodes in identification of Chinese materia medica for the first time. All genomic DNAs from thirty one samples were extracted. The ITS (internal transcribed spacer) regions were amplified and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. And the sequences of the ITS regions were aligned through Clustal-W and the genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model. The neighbor-joining (NJ) phylogenetic trees were constructed. The ITS2 regions were obtained by using the hidden Markov model (HMM)-based annotation methods from the ITS sequences. Results indicated that the lengths of ITS regions of Notopterygii Rhizoma et Radix were 603-604 bp, while the lengths of ITS2 regions were 228 bp. The haplotypes of ITS/ITS2 regions of Notopterygii Rhizoma et Radix were the same as those of the original plant leaves. The intra-specific genetic distances were smaller than inter-specific ones in ITS/ITS2 regions of Notopterygium incisum and N. franchetii. The NJ trees showed that N. incisum, N. franchetii and its adulterants can be easily differentiated according to their monophyly. Therefore, ITS/ITS2 regions as DNA barcodes can stably and accurately distinguish Notopterygii Rhizoma et Radix from its adulterants and could provide a new technique to ensure clinical safety in utilization of traditional Chinese medicines.

[Identification of Daturae flos and its adulterants based on DNA barcoding technique].

To identify the original plant of Daturae Flos from its adulterants by DNA barcoding, the sequences of ITS2, psbA-trnH, matK, rbcL of four species including Datura metel, Darura innoxia, Darura stramonium and Brugmansia arborea were compared and analyzed. The PCR and sequencing success rate of the four regions (ITS2, psbA-trnH, matK, rbcL) was 100%, 90%, 100% and 85%, respectively. Sequences were assembled with CodonCode Aligner. K2P distances were calculated and NJ tree was performed by MEGA 4.1. Thirty SNPs were found among ITS2 sequences, and 33 insert/deletes were found among psbA-trnH intergenic regions. The interspecific K2P distance of ITS2 and psbA-trnH was obviously higher than that of the intraspecific one. As to matK and rbcL, there was no "Barcoding Gap" existing between inter- and intra-specific distances. The NJ trees of the four regions/combinations were built separately. Samples of Brugmansia arborea were clustered into one clade, and the other species of Datura L. formed another clade. The results showed that either ITS2 or psbA-trnH was useful to identify Daturae Flos from its adulterants.

Identification of Cistanche species (Orobanchaceae) based on sequences of the plastid psbA-trnH intergenic region.

The dried succulent stems of Cistanche (Cistanche deserticola Y. C. Ma and Cistanche tubulosa Wight.) are one of the most widely used components of traditional Chinese medicines. However, it is often confused and substituted with the roots of Orobanche pycnostachya, Boschniakia rossica (Cham. & Schltdl.) Standl., Cistanche sinensis Beck, and Cistanche salsa (C. A. Mey.) Beck. In this study, we identified psbA-trnH regions from species and tested their suitable for the identification of the above mentioned taxa. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Cistanche species. Additionally, the average genetic distance of psbA-trnH ranging from 0.077% to 0.743%. In contrast, the intra-specific variation among Cistanche species was found to be significantly different from those of other species, with percentages of variation studied ranged from 0% to 0.007%. The sequence difference between the psbA-trnH sequences of Cistanche species and Orobanche pycnostachya ranged from 0.979% to 1.149%. The distance between the Cistanche species and Boschniakia rossica ranged from 1.066% to 1.224%. Our results suggest that the psbA-trnH intergenic spacer region represent a barcode that can be used to identify Cistanche species and other morphologically undistinguishable species.

Identification of Dendrobium species by a candidate DNA barcode sequence: the chloroplast psbA-trnH intergenic region.

DNA barcoding is a novel technology that uses a standard DNA sequence to facilitate species identification. Although a consensus has not been reached regarding which DNA sequences can be used as the best plant barcodes, the psbA-trnH spacer region has been tested extensively in recent years. In this study, we hypothesize that the psbA-trnH spacer regions are also effective barcodes for Dendrobium species. We have sequenced the chloroplast psbA-trnH intergenic spacers of 17 Dendrobium species to test this hypothesis. The sequences were found to be significantly different from those of other species, with percentages of variation ranging from 0.3 % to 2.3 % and an average of 1.2 %. In contrast, the intraspecific variation among the Dendrobium species studied ranged from 0 % to 0.1 %. The sequence difference between the psbA-trnH sequences of 17 Dendrobium species and one Bulbophyllum odoratissimum ranged from 2.0 % to 3.1 %, with an average of 2.5 %. Our results support the notion that the psbA-trnH intergenic spacer region could be used as a barcode to distinguish various Dendrobium species and to differentiate Dendrobium species from other adulterating species.

Determination and pharmacokinetics of danshensu in rat plasma after oral administration of danshen extract using liquid chromatography/tandem mass spectrometry.

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated for the determination and pharmacokinetics of danshensu in rat plasma samples using ferulic acid as internal standard (IS). The plasma samples were treated by liquid-liquid extraction, and the analyses were determined using electrospray negative ionization mass spectrometry in selected reaction monitoring (SRM) mode. The signal intensity of the m/z 196.8 --> 134.8 transition of danshensu was found to relate linearly to danshensu concentrations in the plasma from 5-500 ng/mL. The lower limit of quantification (LLOQ) as determined by the LC/MS/MS method amounted to 5 ng/mL. The intra- and inter-day precision was below 10.82%, and the accuracy was between -3.51% and +11.92%. The validated LC/MS/MS method was applied to a pharmacokinetic study in which danshen extract (containing 40 mg/g danshensu) was administered orally to rats at a single dose of 200 mg/kg in 2% water.

[The growth of Salvia miltiorrhiza seedling and root system following transplantation].

OBJECTIVE: To study the relationship between plant growth and accumulation of active ingredients in Salvia miltiorrhiza. METHOD: Transplants of S. miltiorrhiza were sampled at 20 day intervals. At each stage, the growth of seedling and root system was recorded and the contents of tanshinone II (A) and salvia acid were measured. RESULT AND CONCLUSION: This study showed that the rapid growth stage of the root system lags behind that of the seedling system, but the growing period of root system lasts longer. The quantitative change of roots reveals a double "S" curve; two rapid growth stages emerge during 30 - 70 days and 140 - 200 days after the seedlings were transplanted. The content of salvia acid reaches the highest level during 140 - 180 days, whereas the content of tanshinone peaks during 100 - 120 days.

[Study on the characteristic of assimilating nitrogenous phosphorous fertilizer and the accumulation disciplinarian of total tanshinons of Salvia miltiorrhiza].

OBJECTIVE: To confirm the amount of fertilizer and the ration of fertilizer. METHOD: Through the experiment of planting Salvia miltiorrhiza in pot and in field were carried out. RESULT: When N:P = 1:1, The production was 1.8 times more than the contrast group in plotting experiment, and in field the production of the highest fertilizer plot were 2.5 times more than the contrast and the higher and high fertilizer plot is 2.25 and 1.2 times respective were than those in the contrast group. CONCLUSION: The proper ration of nitrogenous fertilizer and phosphorous fertilizer is 1:1. Nitrogen shows negative effects to the accumulation of tanshinon IIA, the more the nitrogen ous fertilizer, The less the content of tanshinon IIA. In contrast, phosphor ous fertilizer shows good effects on the accumulation of tanshinon IIA. Phosphorous fertilizer could alleviate the decline of the content of tanshinon IIA by using nitrogenous fertilizer. The accumulation peak of the tanshinon IIA emerge in the period of 150 d.

[The short-term and long-term treatment outcomes in patients with pulmonary tuberculosis positive for drug-resistant and sensitive strains].

OBJECTIVES: To analyze the treatment outcomes in patients with smear positive tuberculosis, and to compare the difference in treatment response among patients infected with drug-sensitive and drug-resistant strains. METHODS: From 1998 to 2000, seven hundred and seventy-seven patients with primary smear-positive tuberculosis, which were from 30 surveillance sites, were followed for two years to monitor their treatment outcomes. RESULTS: At the completion of the 6 months' therapy, the overall rate of treatment failure was 1.8%, 2.6% for the drug-resistant cases and 1.6% for the drug-sensitive cases. Six-month follow-up showed a positive conversion rate of 2.7% in all the cases, 8.5% and 1.2% (P < 0.005) in the drug-resistant and the drug-sensitive cases respectively. One year follow-up showed that the positive conversion rate was 2.6% in all the cases, 6.9% and 1.6% (P < 0.005) in the drug-resistant and the drug-sensitive cases, respectively. Two-year follow-up showed an overall positive conversion rate of 1.3%, 1.0% and 1.3% in the drug-resistant and the drug-sensitive cases, respectively. Of the 152 drug-resistant cases, the rate of treatment failure was 2.6% at the completion of 6 months' therapy, but in cases with MDR-TB the rate was 10.3%. Six-month follow-up showed an overall positive conversion rate of 8.5%, but the rate reached 37.0% in cases with MDR-TB. One-year and two-year follow-up showed that the positive conversion rates were 6.9% and 1.0% respectively in all the drug-resistant cases, but 6.3% and 6.7% respectively in the MDR cases. CONCLUSIONS: Under the guidelines of the National Tuberculosis Program (NTP), the 2H(3)R(3)S(3)Z(3)/4H(3)R(3) regimen for primary smear-positive pulmonary TB was effective. But the cure rate was lower and the positive conversion rate higher in patients with MDR-TB.