Chloroplast Genome Sequences and Phylogenetic Analysis of Eight Newly Sequenced Caryophyllaceae Species.

BACKGROUND: Caryophyllaceae is a big family composed of many economic and medicinal species. However, the phylogeny of the family is insufficient and genome data are lacking for many species. OBJECTIVE: Using next-generation sequencing (NGS) to acquire the chloroplast (cp) genomes of Eremogone acicularis (F.N.Williams) Ikonn., E. brevipetala (Tsui & L.H.Zhou) Sadeghian & Zarre, E. bryophylla (Fernald) Pusalkar & D.K.Singh, E. kansuensis (Maxim.) Dillenb. & Kadereit, Shivparvatia glanduligera (Edgew.) Pusalkar & D.K.Singh, Silene atsaensis (Marq.) Bocquet, S. caespitella Williams, and S. lhassana (Williams) Majumdar. METHODS: Bioinformatic software was used to conduct the comparative genome and phylogeny analysis of these cp genomes. RESULTS: The eight cp genomes were 132 188-151 919 bp in length, containing 130-132 genes. A/T was dominant in simple sequence repeats (SSRs). Forward repeats and palindromic repeats were the most frequent in long terminal repeats (LTRs). Compared with the four species of Eremogone Fenzl, the inverted repeat (IR) boundaries of S. caespitella, S. atsaensis, S. lhassana, and Sh. glanduligera were significantly expanded. Four and one mutational hotspots were identified in the large single copy (LSC) region and small single copy (SSC) region, respectively. The ratio of nonsynonymous substitution to synonymous substitution (Ka/Ks ratio) showed these cp genomes may have undergone strong purifying selection. In the phylogenetic trees, both Silene L. and Eremogone were monophyletic groups. However, Sh. glanduligera was closely related to Amaranthus hypochondriacus. CONCLUSION: These results have provided new evidence and useful information for species identification, evolution, and genetic research on the Caryophyllaceae. HIGHLIGHTS: In this study, eight newly sequenced cp genomes of Caryophyllaceae species were reported for the first time.

Comparative analyses of five complete chloroplast genomes from the endemic genus Cremanthodium (Asteraceae) in Himalayan and adjacent areas.

Cremanthodium Benth. is an endemic genus in the Himalayas and adjacent areas. Some plants of the genus are traditional medicinal plants in Tibetan medicine. In this study, the chloroplast genomes of five species (Cremanthodium arnicoides (DC. ex Royle) Good, Cremanthodium brunneopilosum S. W. Liu, Cremanthodium ellisii (Hook. f.) Kitam., Cremanthodium nervosum S. W. Liu, and Cremanthodium rhodocephalum Diels) were collected for sequencing. The sequencing results showed that the size of the chloroplast genome ranged from 150,985 to 151,284 bp and possessed a typical quadripartite structure containing one large single copy (LSC) region (83,326-83,369 bp), one small single copy (SSC) region (17,956-18,201 bp), and a pair of inverted repeats (IR) regions (24,830-24,855 bp) in C. arnicoides, C. brunneopilosum, C. ellisii, C. nervosum, and C. rhodocephalum. The chloroplast genomes encoded an equal number of genes, of which 88 were protein-coding genes, 37 were transfer ribonucleic acid genes, and eight were ribosomal ribonucleic acid genes, and were highly similar in overall size, genome structure, gene content, and order. In comparison with other species in the Asteraceae family, their chloroplast genomes share similarities but show some structural variations. There was no obvious expansion or contraction in the LSC, SSC or IR regions among the five species, indicating that the chloroplast gene structure of the genus was highly conserved. Collinearity analysis showed that there was no gene rearrangement. The results of the phylogenetic tree showed that the whole chloroplast genomes of the five species were closely related, and the plants of this genus were grouped into one large cluster with Ligularia Cass. and Farfugium Lindl.

Comparative and Phylogenetic Analysis of Chloroplast Genomes of Two Medicinal Species of Spatholobus.

BACKGROUND: The vine stem of Spatholobus suberectus Dunn (S. suberectus), called "JiXueTeng", has been used as a significant medicine for thousands of years in China. However, reliable field identification of this medicinal plant remains problematic, inaccurate identification may cause serious adverse effects in the functions of the drug and may affect the clinical medication reviews. OBJECTIVE: To ensure use of the exact medicine and implement protective legislation, it is imperative to obtain the chloroplast (cp) genome of S. suberectus, which can be used as a valuable resource for species identification and phylogenetic analysis. METHODS: In this study, the complete cp genomes of S. suberectus (152 173 bp (base pair)) and S. pulcher (151 099 bp) were assembled for the first time by using next-generation sequencing (NGS) technology to gain abundant information on the genus of Spatholobus. And some bioinformatics softwares were used for data filtering, assembling and analyzing. RESULTS: We found the G and C contents of S. suberectus and S. pulcher were close, 35.19% and 35.37%, respectively. The noncoding regions were more divergent than coding ones. Moreover, we revealed eight divergence hotspots (trnH, trnK-rbcL, trnL-rbcT, psbD-trnT, trnC-rpoB, atpI-atpH, ycf4, and trnL-rpl32) which might be used as candidate molecular markers for Spatholobus identification. The analysis of the phylogenetic relationship indicated that two Spatholobus species were clustered together and two Spatholobus species was sister to the Cajanus. CONCLUSION: The findings of this study were conducive to species identification and phylogenetic research of Spatholobus and provided valuable resources for finding the substitution of S. suberectus. HIGHLIGHTS: We assembled the complete cp genomes of S. suberectus and S. pulcher for the first by using next-generation sequencing.

[Activity of Codonopsis canescens against rheumatoid arthritis based on TLRs/MAPKs/NF-κB signaling pathways and its mechanism].

This paper aims to explore the activity of Codonopsis canescens extract against rheumatoid arthritis(RA) based on the Toll-like receptors(TLRs)/mitogen-activated protein kinases(MAPKs)/nuclear factor kappa B(NF-κB) signaling pathways and its mechanism. The ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry(UPLC-Q-TOF-MS) was used to identify the components of C. canescens extract. Forty-eight male SD rats were randomly divided into six groups, namely the normal group, the model group, the methotrexate(MTX) tablet group, and the low, medium, and high-dose C. canescens extract(ZDS-L, ZDS-M, and ZDS-H) groups, with 8 rats in each group. The model of collagen-induced arthritis in rats was induced by injection of bovine type Ⅱ collagen emulsion. MTX(2.5 mg·kg~(-1)), ZDS-L, ZDS-M, and ZDS-H(0.3 g·kg~(-1), 0.6 g·kg~(-1), and 1.2 g·kg~(-1)) were administrated by gavage. Rats in the normal group and the model group received distilled water. MTX was given once every three days for 28 days, and the rest medicines were given once daily for 28 days. Body weight, degree of foot swelling, arthritis index, immune organ index, synovial histopathological changes, and serum levels of tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß), and interleukin-6(IL-6) were observed. Protein expressions of TLR2, TLR4, NF-κB p65, p38 MAPK, and p-p38 MAPK in rats were determined by Western blot. Thirty-four main components were identified by UPLC-Q-TOF-MS, including 15 flavonoids, 7 phenylpropanoids, 4 terpenoids, 4 organic acids, 2 esters, and 2 polyalkynes. As compared with the normal group, the body weight of the model group was significantly decreased(P<0.01), and foot swelling(P<0.05, P<0.01), arthritis index(P<0.01), and the immune organ index(P<0.01) were significantly increased. The synovial histopathological injury was obviously observed in the model group. The serum levels of inflammatory factors TNF-α, IL-1ß, and IL-6 were significantly increased(P<0.01), and the protein expression levels of TLR2, TLR4, NF-κB p65, p-p38 MAPK/p38 MAPK in the synovial tissue were significantly increased(P<0.01) in the model group. As compared with the model group, the body weights of the ZDS dose groups were increased(P<0.01), and the degree of foot swelling(P<0.01) and the arthritis index were decreased(P<0.05, P<0.01). The immune organ index was decreased(P<0.01) in the ZDS dose groups, and the synovial tissue hyperplasia and inflammatory cell infiltration were alleviated. The serum levels of TNF-α, IL-1ß, and IL-6 were significantly decreased(P<0.05, P<0.01), and the protein expression levels of TLR2, TLR4, NF-κB p65, p-p38 MAPK/p38 MAPK were decreased(P<0.05, P<0.01) in the ZDS dose groups. C. canescens extract containing apigenin, tricin, chlorogenic acid, aesculin, ferulic acid, caffeic acid, and oleanolic acid has a good anti-RA effect, and the mechanism may be related to the inhibition of TLRs/MAPKs/NF-κB signaling pathways.

Lipskynoids A-G, New Acyclic Diterpenes from the Flowers of Carpesium lipskyi.

Seven new acyclic diterpenes, namely lipskynoids A-G (1-7), were isolated from the flowers of Carpesium lipskyi, a traditional Tibetan herbal medicine with anti-inflammatory and antipyretic-analgesic effects. These new compounds were elucidated by analysis of extensive spectroscopic data including ESI-MS, 1D, 2D NMR, and DP4+ analyses. Biological assays showed that 1-7 display significant inhibitory effects against the NO production in LPS-induced RAW264.7 cells with its IC50 values from 9.9 to 18.47 µM, however, no cytotoxicity effect was observed of these isolates against the growth of HePG2, PC3, DU145, and A549 cells.

Carpelipines C and D, Two Anti-Inflammatory Germacranolides from the Flowers of Carpesium lipskyi Winkl. (Asteraceae).

Two new germacranolides, carpelipine C (1) and carpelipine D (2), together with four known ones (3-6), were isolated from Carpesium lipskyi Winkl. flowers, a folk Tibetan herbal medicine with antipyretic-analgesic and anti-inflammatory effects. The chemical structures of new structure were illuminated by diversified spectroscopic and X-ray crystallographic analyses. Compounds 1 and 3 dramatically suppressed the synthesis of NO and decreased pre-inflammatory protein expression of iNOS and COX-2 in LPS-induced RAW264.7 cells. Furthermore, it was revealed that NF-κB/MAPK signaling pathway were involved in the anti-inflammatory process of 1 and 3, and their effects on reducing oxidative stress by activating Nrf2/HO-1 pathway were also measured. This article indicated that the traditional use of C. lipskyi to treat inflammatory diseases has a certain rationality.

Two Olean-27-carboxylic Acid-Type Triterpenoids as Chemical Markers Isolated from Saxifraga umbellulata.

Two olean-27-carboxylic acid-type triterpenoids (1 and 2) were isolated from Saxifraga umbellulata (Saxifragaceae), representing the first case in the chemical discoveries of genus Saxifraga. Compound 1 was determined to be a new compound named 'Saxifragic acid' based on the comprehensive spectroscopic and X-ray crystallographic analyses. Compound 2 (deacetylated saxifragic acid) is a known compound reported before, but its absolute configuration through X-ray crystallographic analyses was first described here. In addition, their cytotoxicity against five digestive human cancer cell lines (BGC-823, GBC-SD, CCC-9810, HT-29, and HepG2) and hepatoprotective activity against CCl4 -induced L-o2 cell injury in vitro were evaluated. Interestingly, UPLC-QTOFMS analysis showed that these two compounds could be used as chemical markers to discriminate between S. umbellulata and S. tangutica, both of which are used for the treatment of hepatobiliary diseases in traditional Tibetan medicine.

Highland barley Monascus purpureus Went extract ameliorates high-fat, high-fructose, high-cholesterol diet induced nonalcoholic fatty liver disease by regulating lipid metabolism in golden hamsters.

ETHNOPHARMACOLOGICAL RELEVANCE: Hepatocyte lipid accumulation is the main feature in the early stage of nonalcoholic fatty liver disease (NAFLD). Highland barley Monascus purpureus Went (HBMPW), a fermentation product of Hordeum vulgare Linn. var. nudum Hook. f. has traditionally been used as fermented foods in Tibet with the effect of reducing blood lipid in folk medicine. AIM OF THE STUDY: This study investigated the protective effects and molecular mechanism of highland barley Monascus purpureus Went extract (HBMPWE) on NAFLD in syrian golden hamster fed with high-fat, high-fructose, high-cholesterol diet (HFFCD). MATERIALS AND METHODS: HFFCD-induced NAFLD golden hamster model was established and treated with HBMPWE. Liver index, biochemical index, and hematoxylin and eosin (HE) staining were observed. Liver metabolomics and western blot analysis were employed. RESULTS: Our study found that HBMPWE ameliorated HFFCD induced dyslipidemia, weight gain and elevated the liver index. In addition, HBMPWE treatment significantly attenuated lipid accumulation in the liver and modulated lipid metabolism (sphingolipid, glycerophospholipid). Our data demonstrated that HBMPWE not only regulated the expression of proteins related to fatty acid synthesis and decomposition (SREBP-1/ACC/FAS/AceS1, PPARα/ACSL/CPT1/ACOX1), but also regulated the expression of proteins related to cholesterol synthesis and clearance (HMGCR, LDLR, CYP7A1). CONCLUSIONS: HBMPWE improved NAFLD through multiple pathways and multiple targets in body metabolism and could be used as a functional food to treat NAFLD and other lipid metabolic disorders.

Two-photon calcium imaging of neuronal and astrocytic responses: the influence of electrical stimulus parameters and calcium signaling mechanisms.

Objective. Electrical brain stimulation has been used to ameliorate symptoms associated with neurologic and psychiatric disorders. The astrocytic activation and its interaction with neurons may contribute to the therapeutic effects of electrical stimulation. However, how the astrocytic activity is affected by electrical stimulation and its calcium signaling mechanisms remain largely unknown. This study is to explore the influence of electrical stimulus parameters on cellular calcium responses and corresponding calcium signaling mechanisms, with a focus on the heretofore largely overlooked astrocytes.Approach. Usingin vivotwo-photon microscopy in mouse somatosensory cortex, the calcium activity in neurons and astrocytes were recorded.Main results. The cathodal stimulation evoked larger responses in both neurons and astrocytes than anodal stimulation. Both neuronal and astrocytic response profiles exhibited the unimodal frequency dependency, the astrocytes prefer higher frequency stimulation than neurons. Astrocytes need longer pulse width and higher current intensity than neurons to activate. Compared to neurons, the astrocytes were not capable of keeping sustained calcium elevation during prolonged electrical stimulation. The neuronal Ca2+influx involves postsynaptic effects and direct depolarization. The Ca2+surge of astrocytes has a neuronal origin, the noradrenergic and glutamatergic signaling act synergistically to induce astrocytic activity.Significance. The astrocytic activity can be regulated by manipulating stimulus parameters and its calcium activation should be fully considered when interpreting the mechanisms of action of electrical neuromodulation. This study brings considerable benefits in the application of electrical stimulation and provides useful insights into cortical signal transduction, which contributes to the understanding of mechanisms underlying the therapeutic efficacy of electrical stimulation for neurorehabilitation applications.

The complete chloroplast genome of Mitreola yangchunensis (Loganiaceae).

The complete chloroplast (cp) genome of Mitreola yangchunensis was sequenced and assembled for the first time. The genome is 154,665 bp in length, containing a large single-copy (LSC) region of 85,351 bp, a small single-copy region (SSC) of 18,218 bp and a pair of inverted repeats (IRs) of 25,548 bp. It contains 113 unique genes, including 79 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. The overall GC content is 37.9%, while the corresponding values of LSC, SSC, and IR regions are 35.9, 32.0, and 43.4%, respectively. Phylogenetic analyses using complete cp genomes showed that M. yangchunensis is most closely related to Mitrasacme pygmaea in Loganiaceae, and Gelsemiaceae and Loganiaceae form a single cluster with high support value.

The complete chloroplast genome of Stephania tetrandra (Menispermaceae).

The complete chloroplast genome of Stephania tetrandra was sequenced and assembled for the first time. The chloroplast genome is 159,974 bp in length, containing a large single-copy (LSC) region of 90,539 bp and a small single-copy region (SSC) of 20,735 bp, separated by a pair of inverted repeats (IRs) of 24,350 bp. The genome contains 113 unique genes, including 79 protein-coding genes (PCGs), 30 tRNA genes, and four rRNA genes. Among them, 15 genes have one intron each and three genes contain two introns. The overall GC content is 37.8%, while the corresponding values of LSC, SSC, and IR regions are 35.8, 32.4, and 43.7%, respectively. Phylogenetic analysis showed that S. tetrandra is more closely related to the clade of two species within Stephania, providing new insight into the evolution of Menispermaceae.

The complete chloroplast genome of Mitrasacme pygmaea (Loganiaceae).

The complete chloroplast genome of Mitrasacme pygmaea was sequenced and assembled for the first time. The chloroplast genome is 152,611 bp in length, containing a large single-copy (LSC) region of 83,881 bp and a small single-copy region (SSC) of 18,110 bp, separated by a pair of inverted repeats (IRs) of 25,310 bp. The genome contains 113 unique genes, including 79 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Among them, 15 genes have one intron each and 3 genes contain two introns. The overall GC content is 37.9%, while the corresponding values of LSC, SSC, and IR regions are 36.0%, 31.7%, and 43.4%, respectively. Phylogenetic analysis showed that M. pygmaea is sister to Gentiana tibetica and provided new insight into the evolution of Loganiaceae.

The complete chloroplast genome of Turpinia arguta (Staphyleaceae).

The complete chloroplast genome of Turpinia arguta was sequenced and assembled for the first time. The chloroplast genome was 160,139 bp in length, containing a large single-copy region (LSC) of 89,625 bp and a small single-copy region (SSC) of 18,262 bp, separated by a pair of inverted repeats (IRs) of 26,126 bp. The genome contained 113 unique genes, including 79 protein-coding genes, 30 tRNA genes, and four rRNA genes. Among them, 15 genes had one intron each and 3 genes containing two introns. The overall GC content was 37.4%, while the corresponding values of LSC, SSC, and IR regions were 35.4%, 31.8%, and 42.8%, respectively. Phylogenetic analysis showed that T. arguta is more closely related to Staphylea trifolia and provided new insight into the evolution of Staphyleaceae.

Cortical Plasticity Induced by Anodal Transcranial Pulsed Current Stimulation Investigated by Combining Two-Photon Imaging and Electrophysiological Recording.

Anodal-transcranial pulsed current stimulation (a-tPCS) has been used in human studies to modulate cortical excitability or improve behavioral performance in recent years. Multiple studies show crucial roles of astrocytes in cortical plasticity. The calcium activity in astrocytes could regulate synaptic transmission and synaptic plasticity. Whether the astrocytic activity is involved in a-tPCS-induced cortical plasticity is presently unknown. The purpose of this study is to investigate the calcium responses in neurons and astrocytes evoked by a-tPCS with different current intensities, and thereby provides some indication of the mechanisms underlying a-tPCS-induced cortical plasticity. Two-photon calcium imaging was used to record the calcium responses of neurons and astrocytes in mouse somatosensory cortex. Local field potential (LFP) evoked by sensory stimulation was used to assess the effects of a-tPCS on plasticity. We found that long-duration a-tPCS with high-intensity current could evoke large-amplitude calcium responses in both neurons and astrocytes, whereas long-duration a-tPCS with low-intensity current evoked large-amplitude calcium responses only in astrocytes. The astrocytic Ca2+ elevations are driven by noradrenergic-dependent activation of the alpha-1 adrenergic receptors (A1ARs), while the intense Ca2+ responses of neurons are driven by action potentials. LFP recordings demonstrated that low-intensity a-tPCS led to enhancement of cortical excitability while high-intensity a-tPCS resulted in diminution of cortical excitability. The results provide some evidence that the enhancement of a-tPCS-induced cortical excitability might be partly associated with calcium elevation in astrocytes, whereas the diminution of a-tPCS-induced cortical excitability might be caused by excessive calcium activity in neurons. These findings indicate that the appropriate current intensity should be used in the application of a-tPCS.

Syringa pinnatifolia Hemsl. fraction protects against myocardial ischemic injury by targeting the p53-mediated apoptosis pathway.

BACKGROUND: Peeled stems of Syringa pinnatifolia Hemsl. (SP) have been widely used to treat extra "He-Yi" induced myocardial ischemia for hundreds of years in Inner Mongolia, China and previous result showed that intragastric pretreatment with total extract (T) of SP has a protective effect against myocardial infarction (MI). HYPOTHESIS: This study aims to describe the pharmacological investigation and chemical characterization of the major (M) and minor (N) fractions obtained from T through column chromatography fractionation on macroporous resin and to explore whether the regulatory effects were linked to the p53-mediated apoptosis pathways. STUDY DESIGN: Left anterior descending (LAD) coronary artery-ligated mice and H9c2 cells cultured in serum-free medium under hypoxic conditions were treated with T, M, and N. METHODS: Echocardiography was performed and biomarkers in serum were determined in mice, and pathological changes were observed through histopathology assay. Immunofluorescence staining and qRT-PCR were used to detect the expression levels of p53 in heart tissue. Flow cytometry was used to measure the level of apoptosis and caspase-3 activity in H9c2 cells. Western blot analysis was conducted to detect p53 and p53-mediated proteins apoptosis pathways of in both tissue and H9c2 cells. RESULTS: Both T and M have an equivalent cardioprotective effect whereas N is non-active. M decreased MI-induced myocardial compensatory expansion by decrease of left ventricular end-systolic diameter (LVESd) and left ventricular end-diastolic diameter (LVEDd) and prevented decreases in ejection fraction (EF) and fractional shortening (FS). The MI-induced increased levels of creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), and hypersensitive C-reactive protein (hs-CRP) were decreased and the expanded infarction size was reduced. M could also improve cell viability and inhibit apoptosis in H9c2 cells under hypoxic conditions. Immunofluorescence and qRT-PCR assay showed that M suppressed p53 expression in the myocardium. Western blot analysis showed that M could prevent MI-induced activation of p53-mediated apoptosis pathway in both myocardium and H9c2 cells. CONCLUSION: The results demonstrated that M may protect against myocardial ischemia by improving cardiac function and inhibiting cardiomyocytes apoptosis. Overall, the present findings supported the clinical application of SP and enriched the research of anti-myocardial ischemia drug from traditional medicines.

Evaluation and Comparison of the Inhibition Effect of Astragaloside IV and Aglycone Cycloastragenol on Various UDP-Glucuronosyltransferase (UGT) Isoforms.

As one of the main active ingredients from Radix Astragali (RA), orally dosed astragaloside IV (AST) is easily transformed to sapogenin-cycloastragenol (CAG) by deglycosylation in the gastrointestinal tract. Because the potential adverse effects of AST and CAG remain unclear, the present study in this article was carried out to investigate the inhibition effects of AST and CAG on UDP-glucuronosyltransferases (UGTs) to explore potential clinical toxicity. An in vitro UGTs incubation mixture was employed to study the inhibition of AST and CAG towards UGT isoforms. Concentrations of 100 µM for each compound were used to initially screen the inhibitory efficiency. Deglycosylation of AST to CAG could strongly increase the inhibitory effects towards almost all of the tested UGT isoforms, with an IC50 of 0.84 µM and 11.28 µM for UGT1A8 and UGT2B7, respectively. Ulteriorly, the inhibition type and kinetics of CAG towards UGT1A8 and UGT2B7 were evaluated depending on the initial screening results. Data fitting using Dixon and Lineweaver-Burk plots demonstrated that CAG competitively inhibited UGT1A8 and noncompetitively inhibited UGT2B7. From the second plot drawn with the slopes from the Lineweaver-Burk plot versus the concentrations of CAG, the inhibition constant (Ki) was calculated to be 0.034 µM and 20.98 µM for the inhibition of UGT1A8 and UGT2B7, respectively. Based on the [I]/Ki standard ([I]/Ki < 0.1, low possibility; 1 > [I]/Ki > 0.1, medium possibility; [I]/Ki > 1, high possibility), it was successfully predicted here that an in vivo herb-drug interaction between AST/CAG and drugs mainly undergoing UGT1A8- or UGT2B7-catalyzed metabolism might occur when the plasma concentration of CAG is above 0.034 µM and 20.98 µM, respectively.

[Analysis of varieties and standards of Scrophulariaceae plants used in Tibetan medicine].

In this paper, the popular domestic varieties and quality standard of Scrophulariaceae plants used in Tibetan medicine were analyzed. The results showed that there were 11 genera and 99 species (including varieties), as well as 28 medicinal materials varieties of Scrophulariaceae plants were recorded in the relevant literatures. In relevant Tibetan standards arid literatures, there are great differences in varieties, sources, parts, and efficacies of medicinal plant. Among them, about 41.4% (including 41 species) of endemic plants, about 15.2% (including 15 species) of the original plants have medicinal standard legal records, except the medicinal materials of Scrophalaria ningpoensis, Lagotis brevituba, Picrorhiza scrophulariiflora, Veronica eriogyne general, most varieties have not completed quality standard. Consequently it is necessary to reinforce the herbal textual, resources and the use present situation investigation, the effects of the species resources material foundation and biological activity, quality standard, specification the medical terms of the plants, and promote Tibetan medicinal vareties-terminologies-sources such as the criterion and quality standard system for enriching the varieties of Tibetan medicinal materials and Chinese medicinal resources.

[Traightened on Chinese endemic seed plant species of medicine plants used in Tibetan medicine].

This paper is in order to discussion with the composition and characteristics of Tibetan medicine plant resources, and promote the reasonable protection and utilization of the resources of Tibetan materia medica. Statistical analysis of species, distributions, and others of Chinese endemic seed plant from Tibetan medicine plants and usually used in the clinic of Tibetan medicine. The results showed that there are 523 species (25%) of Chinese endemic seed plant, belonging to 65 families and 162 genera, in about 2 000 varieties of Tibetan medicine plants recorded in relevant literatures. There are 180 Chinese endemic seed plant species (28%) belonging to 42 families and 72 genera from 625 medicine plants usually used in the clinic of Tibetan medicine. Specifically, the most of these Chinese endemic seed plant species are characteristic crude drug used in Tibetan medicine, and mainly or only distributed in Qinghai-Tibet Plateau. And a few species of them were intersected with traditional Chinese medicines (TCM) and other ethnic medicines. In addition, about 10% are listed in China Species Red List. The Qinghai-Tibet Plateau is the most abundant areas of Areal-types of the Chinese endemic seed plant. This is the biological and ecological reason formation the characteristics of Tibetan medicine plant resources. Therefore, strengthen the research of Chinese endemic seed plants used in Tibetan medicine is great significance for the reasonable protection and utilization of Tibetan medicine plant resources.

[Analysis of varieties and standards of Leguminosae plants used in Tibetan medicine].

In this paper, the domestic varieties and quality standard of Leguminosae medicinal plants used in Tibetan medicine were analyzed. The results showed that there were 36 genera and 142 species (including varieties), as well as 64 medicinal materials varieties of Leguminosae plants were recorded in relevant literatures. In relevant Tibetan standards and literatures, there are great differences in varieties, sources, used parts, and efficacy of medicinal plants. Among them, about 38.0% (including 54 species) of the endemic plants, about 25.4% (including 36 species) of the original plants have medicinal standard legal records, except 9 kinds of traditional Chinese medicine general quality standard more fairly completed, the most varieties have only description about characters, identification, etc. Therefore it is necessary to reinforce study for the herbal textual, resources and the use present situation, chemical components and biological activity, quality standard, medicinal terms specification, to promote establishment of quality standard system for variety-terminologies-sources of Tibetan medicinal plants.

Simple LC-MS/MS methods for simultaneous determination of pitavastatin and its lactone metabolite in human plasma and urine involving a procedure for inhibiting the conversion of pitavastatin lactone to pitavastatin in plasma and its application to a pharmacokinetic study.

Sometimes, drugs and their metabolites in plasma may convert to each other. This phenomenon is called interconversion, which may result in the instability problem of the plasma samples. The instability problem caused by interconversion of the co-existing metabolites may often be ignored, since there is no drug metabolite in the quality control samples prepared for method validation. Pitavastatin lactone (Pi-LAC), a main metabolite of pitavastatin (Pi), is very unstable and easily converted to Pi in plasma. In this paper, simple and rapid LC-ESI-MS/MS methods were developed for the simultaneous determination of Pi and Pi-LAC in human plasma and urine. The sample stability was examined under different conditions. The interconversion of Pi and Pi-LAC was prevented by adding a pH 4.2 buffer solution to the freshly collected plasma samples. Detection was performed using an electrospray ionization (ESI) operating in positive ion multiple reaction monitoring mode by monitoring the ion transitions from m/z 422.2→290.3 (Pi), 404.2→290.3 (Pi-LAC) and m/z 611.3→423.2 (candesartan cilextetil, the internal standard), respectively. The calibration curve of Pi and Pi-LAC in both human plasma and urine showed good linearity over the concentration range of 0.1-200 ng/mL. The established methods were successfully applied to a pharmacokinetic study of pitavastatin calcium tablets in healthy Chinese volunteers after oral administration of 1, 2 and 4 mg single and multiple doses of pitavastatin calcium. The pharmacokinetic parameters of Pi and Pi-LAC in Chinese volunteers were given respectively. The urinary excretion profiles of Pi and Pi-LAC in Chinese volunteers were also presented. After receiving a single 4 mg oral dose of pitavastatin calcium, the average cumulative urinary excretion percentages of Pi and Pi-LAC in Chinese volunteers were (0.41 ± 0.16)% and (6.1 ± 5.0)%, respectively.