[Qualitative and semiquantitative analyses of the chemical components of the seed coat and kernel of Ziziphi Spinosae Semen].

Ziziphi Spinosae Semen refers to the dried seed of Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou. The seed is composed of a reddish brown coat and a yellow kernel. A comparative study was conducted to investigate differences in the chemical composition and their relative contents between the seed coat and kernel of Ziziphi Spinosae Semen. First, the chemical compounds found in the seed coat and kernel were characterized and identified using ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The analytical results tentatively identified 57 chemical compounds based on reference-compound comparison, literature retrieval, and chemical-database (e. g., MassBank) searches; these compounds included 14 triterpenes, 23 flavonoids, 7 alkaloids, 6 carboxylic acids, and 7 other types of compounds. The mass error of the identified compounds was within the mass deviation range of 5×10-6 (5 ppm). Next, two methods of multivariate statistical analysis, namely, principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), were used to compare the differential compounds between the two seed parts. A total of 17 differential compounds were screened out via OPLS-DA based on a variable importance in projection (VIP) value of >5. The results revealed that betulinic acid, betulonic acid, alphitolic acid, and jujuboside Ⅰ mainly existed in the seed coat whereas the 13 other compounds, such as spinosin, jujuboside A, and 6‴-feruloylspinosin, mainly existed in the seed kernel. Therefore, these 17 differential compounds can be used to distinguish between the two seed parts. Finally, a semiquantitative method was established using UPLC and a charged aerosol detector (CAD) with inverse gradient compensation in the mobile phase. Six representative compounds with different types were selected to examine the CAD response consistency: magnoflorine (alkaloid), spinosin (flavone), 6‴-feruloylspinosin (flavone), jujuboside A (triterpenoid saponin), jujuboside B (triterpenoid saponin), and betulinic acid (triterpenoid acid). The results showed that the relative standard deviation (RSD) of the average response factors at different levels of these six compounds was 7.04% and that their response intensities were similar. Moreover, each compound in the fingerprint demonstrated good response consistency, and the peak areas obtained directly reflected the contents of each compound. Based on the semiquantitative fingerprints obtained, betulinic acid and oleic acid were considered the main components of the seed coat. The betulinic acid content in the seed coat was approximately 7 times higher than that in the seed kernel. Spinosin, jujuboside A, linoleic acid, betulinic acid, and oleic acid were the main components of the seed kernel. The spinosin content in the seed kernel was 18 times higher than that in the seed coat. In addition, the jujuboside A content in the seed kernel was 24 times higher than that in the seed coat. The proposed method can accurately determine the main components and compare the relative contents of these components in different seed parts. In summary, this study identified the differences in chemical components between the seed coat and kernel of Ziziphi Spinosae Semen and clarified the main components and their relative contents in these parts. The findings can not only provide a basis for the identification of chemical compounds and quality research on different parts of Ziziphi Spinosae Semen but also promote the development and utilization of this traditional Chinese medicine.

Dammarane-type saponins from Gynostemma pentaphyllum and their anti-aging activities via up-regulating mitochondria related proteins.

The importance of mitochondria in regulation of aging has been extensively recognized and confirmed. Gynostemma pentaphyllum (Thunb.) Makino, a homology of medicine and food, has been widely utilized as dietary supplement. In this study, the transcriptome of normal cells (wild type mouse embryo fibroblasts) regulated by the 30% aqueous EtOH extract of G. pentaphyllum was firstly evaluated by RNA sequencing and the results revealed that the G. pentaphyllum could up-regulate the genes involved in oxidative phosphorylation (OXPHOS) and sirtuin (SIRT) signaling pathways, indicating its effect in promoting cell viability might be attributed to the role of improving mitochondrial functions. To further discover the bioactive compounds, sixteen undescribed dammarane-type saponins along with twenty-eight known analogues were isolated from the active extract of G. pentaphyllum. Their structures were elucidated by means of comprehensive analysis of NMR and HRMS spectroscopic data. All isolates were evaluated for the regulatory effects on SIRT3 and translocase of the outer membrane 20 (TOM20), and thirteen of them exhibited satisfactory agonist activities on both SIRT3 and TOM20 at 5 µM. Furthermore, the preliminary structure-activity relationships analysis demonstrated the additional hydroxymethyl and carbonyl groups or less sugar residues in saponins could contribute positively to the up-regulatory effect on SIRT3 and TOM20. These findings encouraged the potential roles of G. pentaphyllum and its bioactive saponins in the development of natural drugs for the treatment of aging-related diseases.

An effective and high-throughput sample preparation method involving demalonylation followed by an ultrahigh-performance liquid chromatography-charged aerosol detector for analyzing gypenoside XLIX and gypenoside A in Gynostemma longipes.

Gypenosides (Gps) are the major bioactive components in Gynostemma species. They include neutral Gps and acidic malonylgypenosides (MGps). MGps are abundant in Gynostemma species and can be transformed into corresponding Gps via extraction, concentration, and drying. If only the Gps were quantified and MGps were ignored, the quality of Gynostemma species would be underestimated. This study aimed to develop a sample preparation method involving demalonylation and ultrahigh-performance liquid chromatography-charged aerosol detector (UHPLC-CAD) analysis to determine the contents of gypenoside XLIX (Gp XLIX) and gypenoside A (Gp A). First, the optimized ultrasonic extraction method was established to extract G. longipes powder ultrasonically. Then, the extracted solution was put into a closed container (centrifuge tube) and heated in a water bath at 95 °C. Then, MGps were converted into corresponding Gps. The proposed preparation method was compared with the other three methods, including water bath reflux heating, alkali hydrolysis, and extraction of heated powder, and was shown to exhibit higher conversion and better convenience. Subsequently, an UHPLC-CAD method was established and validated. Gp XLIX and Gp A showed excellent linear correlations between 15.55 and 248.8 µg/mL and 24.10-385.5 µg/mL, respectively (R2 > 0.999). The limit of detection was 1.40 ng (Gp XLIX) and 2.41 ng (Gp A), and the limit of quantification was 7.77 ng and 14.46 ng, respectively. The relative standard deviation for precision, stability, and repeatability was 0.63-3.15%. The average recovery of Gp XLIX and Gp A was 98.97% and 98.23%, respectively. The established method was applied for determining Gp XLIX and Gp A contents in wild or cultivated G. longipes samples collected from the Qinba Mountains area. The contents of Gp XLIX and Gp A were 5.16-23.02 mg/g and 15.78-54.55 mg/g, respectively. Conclusively, the proposed sample preparation and analysis method could be used for the quality control and evaluation of G. longipes.

New prenylated flavonoid glycosides derived from Epimedium wushanense by ß-glucosidase hydrolysis and their testosterone production-promoting effects.

Six new prenylated flavonoid glycosides, including four new furan-flavonoid glycosides wushepimedoside A-D (1-4) and two new prenyl flavonoid derivatives wushepimedoside E-F (5-6), and one know analog epimedkoreside B (7) were isolated from biotransformation products of the aerial parts of Epimedium wushanense. Their structures were elucidated according to comprehensive analysis of HR-MS and NMR spectroscopic data, and the absolute configurations were assigned using experimental and calculated electronic circular dichroism (ECD) data. The regulatory activity of compounds 1-7 on the production of testosterone in primary rat Leydig cells were investigated, and 4 and 5 exhibited testosterone production-promoting activities. Molecular docking analysis suggested that bioactive compounds 4 and 5 showed the stable binding with 3ß-HSD and 4 also had good affinity with Cyp17A1, which suggested that these compounds may regulate testosterone production through stimulating the expression of the above two key proteins.

[Non-alkaloid constituents of Hymenocallis littoralis].

Perennial herb Hymenocallis littoralis(Amaryllidaceae) boasts anti-tumor, anti-virus, and anti-inflammatory activities. As the representative constituents, alkaloids have attracted much attention, whereas the non-alkaloid constituents have been rarely reported. Therefore, this study investigated the non-alkaloid constituents of H. littoralis and their contribution to the various pharmacological activities of the herb. Thirteen non-alkaloid compounds were isolated from the 95% ethanol extract of dried whole plant of H. littoralis after a series of chromatographic separation steps and spectral analysis, and they were identified as 5,7-dihydroxy-6,8-dimethoxy-2-hydroxymethyl-4H-chromoen-4-one(1), undulatoside A(2),(2S)-7,4'-dihydroxyflavane(3), naringenin(4), 4',7-hydroxy-8-methylflavanone(5), 8-methylnaringenin(6), 8-demethylfarrerol(7), 6-methyl-aromadendrin(8), 4',5,7-trihydroxy-8-methylflavanone(9), syzalterin(10), 6-methylapigenin(11), isoliquiritigenin(12), and undatuside C(13) based on the spectroscopic data analysis. Among them, compound 1 was a new chromone derivative, and compounds 2 and 4-13 were isolated form this plant for the first time.

[Progress in research and development of Xanthoceras sorbifolia].

Xanthoceras sorbifolia, an excellent oil-rich woody species, has high comprehensive economic value in edible, medicinal, and ornamental fields. The chemical composition, pharmacological effect, and quality control of X. sorbifolia were introduced, and its development and application were reviewed in this study. As revealed by the previous research, the main chemical constituents of X. sorbifolia were triterpenoids, flavonoids, fatty acids, phenylpropanoids, steroids, phenolic acids, organic acids, etc. It possesses pharmacological effects, such as neuroprotection, bacteriostasis, anti-oxidation, anti-tumor, anti-inflammation, analgesia, anti-HIV, and anti-coagulation. X. sorbifolia is widely applied in medical, food, chemical industry, and other fields, and deserves in-depth research and development.

New steroidal glycosides from the roots of Asparagus cochinchinensis.

Steroidal saponins were the main active constituents of the traditional medicinal herb Asparagus cochinchinensis. A phytochemical investigation of A. cochinchinensis roots led to the isolation of nine new steroidal glycosides (1-9) and seven known analogues (10-16). Their structures were established by spectroscopic analyses as well as necessary chemical evidence.

Anthraquinone analogues with inhibitory activities against influenza a virus from Polygonatum odoratum.

Three anthraquinone analogues (1-3) were isolated by phytochemical work on EtOAc-soluble ingredients extracted from the roots of Polygonatum odoratum. The structures of all isolates were elucidated by NMR, MS and CD experiments, of which 1 (polygodoquinone A) was identified as a new anthraquinone derivative. Specifically, 1 represents an unusual structure composed of a naphthoquinone derivative linked to an anthraquinone via a C-C bond. 1-3 exhibited remarkable influenza A virus inhibitory activity with IC50 values of 11.4, 11.0, and 2.3 µM, respectively, which were better than ribavirin as the positive control.

Steroidal glycosides, homoisoflavanones and cinnamic acid derivatives from Polygonatum odoratum and their inhibitory effects against influenza A virus.

A phytochemical investigation of Polygonatum odoratum roots led to the isolation of fifteen steroidal glycosides (1-15), three homoisoflavanones (16-18) and four cinnamic acid derivatives (19-22). The structures of all isolated compounds were established mainly by spectroscopic analyses as well as necessary chemical evidence, of which 1-8 (polygodorasides A-G) were identified as new steroidal glycosides. Among the isolates, compounds 7 and 17 showed remarkable in vitro inhibitory effects against influenza A virus with IC50 values of 14.30 and 49.70 µM (positive control ribavirin 28.4 µM).

Steroidal saponins from Trillium tschonoskii rhizomes and their cytotoxicity against HepG2 cells.

A phytochemical study on the rhizomes of Trillium tschonoskii led to the isolation of fourteen new steroidal saponins, trillitschosides S1-S14 (1-14), along with ten known analogues (15-24). Their structures were established mainly by spectroscopic analyses as well as necessary chemical evidence. All isolated compounds were screened for the cytotoxicity against HepG2 cells, and the results demonstrated that only the known compounds 21-24 exhibited the remarkable cytotoxic activity against HepG2 cells which is much better than the positive control of 5-FU.

[Effect of Chinese herbal therapy on T-lymphocytes of IgA nephropathy patients: a clinical observation].

OBJECTIVE: To observe the effect of Chinese herbal therapy on T-lymphocyte subsets in patients with IgA nephropathy (IgAN). METHODS: Totally 36 inpatients and outpatients at Department of Nephropathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, from June 2011 to June 2013 were recruited in the treatment group, while 20 volunteers were recruited as the healthy control group. Patients in the IgAN group only took Chinese herbal decoctions by syndrome typing for 3 months (except those accompanied with hypertension additionally took antihypertensive agents such as angiotensin-converting enzyme inhibitor and/or dihydropyridines calcium antagonist). No intervention was performed in the healthy control group. The values of Th1, Th2, and CD4+ CD25+ Treg, and red blood cell number in urine were detected using flow cytometry before and after treatment. 24 h urine protein was detected using inmmunoturbidimetry. RESULTS: Compared with the healthy control group, the CD4+ CD25+ Treg level obviously decreased in the IgAN group, showing statistical difference (P < 0.01). In the IgAN group, Th1, 24 h urine protein, and urine red blood cell counts were obviously lower after treatment, showing statistical difference when compared with before treatment (all P < 0.05). CONCLUSION: Chinese herbal therapy could reduce urine erythrocyte number and 24 h urine protein of IgAN patients, and down-regulating Th1 expression might be its mechanism.

Three new monoterpene glycosides from the roots of Paeonia lactiflora.

Three new monoterpene glycosides, 2'-O-benzoylpaeoniflorin, albiflorin R2, and albiflorin R3 (1-3) were isolated from the roots of Paeonia lactiflora. Their structures were elucidated on the basis of spectroscopic means including one- and two-dimensional NMR experiments.

[Epidemiologic survey on the prevalence and distribution of childhood functional constipation in the northern areas of China: a population-based study].

OBJECTIVE: Functional constipation (FC) is a common complaint in childhood but the prevalence still remains unclear, the aim of this study was to explore the prevalence of FC in childhood in the northern parts of China. METHODS: A screening program on FC in childhood was carried out in 5 northern cities (Beijing, Tianjin, Shenyang, Changchun, Harbin) of China according to symptoms under the Rome III criteria. Random clustered sampling of the inhabitants was carried out under stratification of cities, geographic zones, schools or nurseries. Sample size of each area was in proportion to the population of the area. The range of age was 4-14. All subjects under study were requested to fill in a questionnaire distributed by the teachers. The screening program was carried out immediately after a section survey was completed. All together, 20 000 questionnaires were distributed with 19 638 retrievals. According to the result of the screening, a small number of patients who met the criteria were further selected to undergo a detailed clinical examination in the hospital including laboratory examination, colonic transit time, defecography or/and barium enema, electromyologram and anorectal manometry to exclude organic disease of the colon. Simultaneously, a detailed questionnaire was requested to fill under the assistance of trained doctors or medical students. Prevalence of FC of the population was adjusted by the rate of correct diagnosis from the detailed study. RESULTS: The adjusted point prevalence of FC in 5 northern cities of China according to Rome III criteria was 4.73% with higher prevalence rate of FC seen in the Beijing area (5.02%) than in other cities (4.82%, 4.76%, 4.27%, 4.40%, with P < 0.001). Male to female ratio was 1.26:1 with majority of the FC fell in the 4 - 6 year olds (5.76%). CONCLUSION: FC appeared a common disorder in childhood in the northern parts of China which called for greater attention. Higher prevalence was noted in the age groups of 4-6.

[Preliminary study on diagnostic standard of functional constipation in children from five cities in North China].

OBJECTIVE: To explore the symptomatic criteria for diagnosis in children with functional constipation(FC) in North China. METHODS: A screening program for FC in children was carried out in 5 northern cities (Beijing, Tianjin, Shenyang, Changchun, Harbin) of China according to symptoms using Rome III( criteria. Random clustered sampling of the inhabitants was carried out under stratification of city, region and school or kindergarten. Sample size of each area was in proportion to the population of the area. The range of age was 4-14. All the subjects were requested to fill in a questionnaire distributed by the teacher. The screening program was carried out immediately after a plot survey. A total of 20,000 questionnaires were distributed and 19,638 retrieved. According to the result of the screening, a small number of patients who fulfilled the criteria were further selected to undergo detail clinical examinations in the hospital including laboratory examination, colonic transit time, defecography or/and barium enema, electromyogram and anorectal manometry to exclude organic disease of the colon. A detailed questionnaire was filled in with the assistance from a trained doctor or a medical student. Potential risk factors and the relative symptoms were explored by comparing frequencies between FC group and non-FC group using χ2 and Logistic analysis. RESULTS: Eighteen symptoms (defecation less than 3 times per week, dry hard sausage-shape stool, difficulty in defecation, abdominal pain, nausea and vomiting, etc) could exist in control group, but FC group had a higher frequency. The difference was statistically significant. Multivariable stepwise logistic regression analysis showed that defecation less than 3 times per week, hard sausage-shape feces, difficulty in defecation, and the need for laxative were associated with FC. The absence of other symptoms had no influence on diagnosis of FC. The sensitivity, specificity and diagnostic accuracy of symptomatic standard diagnostic test based on Rome III( criteria in FC children showed that ≥2 items had the best diagnostic accuracy but moderate sensitivity and specificity. CONCLUSION: The symptomatic criteria for FC diagnosis in children are suggested as follows: (1) defecation frequency less than 3 times per week; (2) dry, hard, sausage-shape stool in most defecation; (3) difficulty in most defecation; (4) use of medication or digital evacuation. FC can be considered when 2 of the above 4 criteria are met and the symptoms last at least 2 months.

Liquiritin potentiate neurite outgrowth induced by nerve growth factor in PC12 cells.

Neurite outgrowth and neuronal differentiation play a crucial role in the development of the nervous system. Understanding of neurotrophins induced neurite outgrowth was important to develop therapeutic strategy for axon regeneration in neurodegenerative diseases as well as after various nerve injuries. It has been reported that extension of neurite and differentiation of sympathetic neuron-like phenotype was modulated by nerve growth factor (NGF) in PC12 cells. In this study, NGF mediated neurite outgrowth was investigated in PC12 cells after liquiritin exposure. Liquiritin is a kind of flavonoids that is extracted from Glycyrrhizae radix, which is frequently used to treat injury or swelling for its life-enhancing properties as well as detoxification in traditional Oriental medicine. The result showed that liquiritin significantly promotes the neurite outgrowth stimulated by NGF in PC12 cells in dose dependant manners whereas the liquiritin alone did not induce neurite outgrowth. Oligo microarray and RT-PCR analysis further clarified that the neurotrophic effect of liquiritin was related to the overexpression of neural related genes such as neurogenin 3, neurofibromatosis 1, notch gene homolog 2, neuromedin U receptor 2 and neurotrophin 5. Thus, liquiritin may be a good candidate for treatment of various neurodegenerative diseases such as Alzheimer's disease or Parkinson's disease.

[Therapeutic effects of curcumin treatment on hepatic fibrosis].

OBJECTIVE: To investigate the therapeutic effects and mechanisms of curcumin treatment on hepatic fibrosis. METHODS: A model of hepatic fibrosis was established using carbon tetrachloride intraperitoneal injections in rats. Curcumin was administered to one group of the model rats (curcumin group) and the other rats were used as controls (control group). Serum levels of ALT, AST, HA, LN, PCIII, and NO were measured, and Hyp, MDA, and SOD in liver tissues were measured. Liver tissue slides were stained with HE and Masson staining to study the pathological changes in the livers. Grades of hepatic fibrosis were evaluated according to a semiquantitative scoring system. RESULTS: In the curcumin group, serum levels of ALT, AST, NO, HA, LN, PCIII, MDA, and Hyp, were (218.50+/-48.89) U/L, (376.60+/-79.13) U/L, (47.96+/-6.53) micromol/L, (289.96+/-60.43) mg/L, (107.35+/-27.24) mg/L, (148.95+/-28.63) microg/L, (236.10+/-30.54) nmol/g, (478.40+/-75.74) microg/g and all were lower than those of the control group (693.75+/-117.57) U/L, (892.50+/-105.69) U/L, (70.95+/-10.23) micromol/L, (468.22+/-93.45) mg/L, (346.44+/-75.08) mg/L, (279.82+/-54.00) microg/L, (402.25+/-39.16) nmol/g, and (752.50+/-77.62) microg/g. The differences were significant. In the curcumin group, the level of SOD (90.39+/-21.23) in the liver tissues was significantly higher than that of the control group (46.52+/-20.01). The hepatic fibrosis scores in the curcumin group were significantly lower than those of the control group. These effects were dose-dependent. CONCLUSIONS: Curcumin reduces rat hepatic fibrosis. Anti-peroxidation and regulation of collagen metabolism in liver tissues may be involved in the therapeutic effectiveness of curcumin on hepatic fibrosis.

Microcalorimetric investigation of the toxic action of Cr(VI) on the metabolism of Tetrahymena thermophila BF(5) during growth.

Tetrahymena thermophila BF(5) produce heat by metabolism and movement. Using a TAM air isothermal microcalorimeter, the power-time curves of the metabolism of T. thermophila BF(5) during growth were obtained and the action on them by the addition of Cr(VI) were studied. The morphological change with Cr(VI) coexisted and biomass change during the process of T. thermophila BF(5) growth were studied by light microscope. Chromium has been regarded as an essential trace element for life. However, hexavalent chromium is a known carcinogen, mutagen, cytotoxicant and strong oxidizing agent. Cr(VI) of different concentration have different effects on T. thermophila BF(5) growth with the phenomenon of low dose stimulation (0-3×10(-5)molL(-1)) and high dose inhibition (3×10(-5) to 2.4×10(-4)molL(-1)). The relationship between the growth rate constant (k) and c is a typical U-shaped curve, which is a characteristic of hormesis. T. thermophila BF(5) cannot grow at all when the concentration of Cr(VI) is up to 2.4×10(-4)molL(-1). The microscopic observations agree well with the results obtained by means of microcalorimetry. And T. thermophila BF(5) had obviously morphological changes by the addition of Cr(VI).