Effects of ginsenoside Rg1-loaded alginate-chitosan microspheres on human bone marrow stromal cells.

The ginsenoside Rg1 is the most abundant compound in ginseng. Recent studies showed that Rg1 had neuroprotective effects on neuronal cells. The present study was to prepare Rg1-loaded alginate-chitosan microspheres and research the effects of microspheres on human bone marrow (BM) stromal cells (hBMSC). The alginate-chitosan microspheres were prepared by mechanical emulsification technique in combination with ion (Ca2+) and chitosan solidification. Subsequently, the microspheres were employed to load Rg1 ginseng extracts. The microspheres had a smooth surface and were spherical in shape. The average diameter of the microspheres was 3.95 µm. The loading efficiency was approximately 2.12%. The purity of isolated hBMSC was over 98.8%. Rg1-loaded microspheres could promote hBMSC proliferation and differentiation. Meanwhile, Rg1-loaded microspheres could also suppress hBMSC apoptosis induced by hypoxia-reoxygenation. In conclusion, these loaded microspheres may be used in the research of neuroprotective effects of Rg1.

Rapidly Simultaneous Determination of Six Effective Components in Cistanche tubulosa by Near Infrared Spectroscopy.

Quantitative determination of multiple effective components in a given plant usually requires a very large amount of authentic natural products. In this study, we proposed a rapid and non-destructive method for the simultaneous determination of echinacoside, verbascoside, mannitol, sucrose, glucose and fructose in Cistanche tubulosa by near infrared spectroscopy (NIRS). Near infrared diffuse reflectance spectroscopy (DRS) and high performance liquid chromatography (HPLC) were conducted on 116 batches of C. tubulosa samples. The DRS data were processed using standard normal variety (SNV) and multiplicative scatter correction (MSC) methods. Partial least squares regression (PLSR) was utilized to build calibration models for components-of-interest in C. tubulosa. All models were then assessed by calculating the root mean square error of calibration (RMSEC), correlation coefficient of calibration (r). The r values of all six calibration models were determined to be greater than 0.94, suggesting each model is reliable. Therefore, the quantitative NIR models reported in this study can be qualified to accurately quantify the contents of six medicinal components in C. tubulosa.

Amelioration of dextran sulphate sodium-induced colitis in mice by echinacoside-enriched extract of Cistanche tubulosa.

Echinacoside (ECH) is a major bioactive phenyethanoids in medicinal herba Cistanche and has been reported to have antiinflammatory activity and beneficial effect on wound healing in many experimental studies. This study was to test the efficacy of ECH-enriched extract of Cistanche tubulosa in the treatment of dextran sulphate sodium (DSS)-induced colitis, a preclinical model of ulcerative colitis. Oral administration of ECH extract significantly suppresses the development of acute colitis, indicated by lowering disease activity index (p < 0.0001, n = 8) and preventing colonic damage (p = 0.0336). Histological examinations showed that ECH extract treatment protected intestinal epithelium from inflammatory injury (p = 0.0249) but had less effect on inflammatory cellular infiltration (p = 0.1753). The beneficial effect of ECH extract treatment was associated with upregulation of transforming growth factor (TGF)-ß1 as well as with an increase in the number of Ki67(+) proliferating cells in diseased colons (p < 0.0001). In cultured MODE-K cells, the addition of ECH extract enhanced in vitro wound healing that depended on TGF-ß1 expression. These data suggest that ECH extract possesses a greater efficacy in preventing DSS-induced colitis in mice, implying the potential of ECH or its derivatives for clinically treating inflammatory bowel disease.

[Effects of Pb2+ stress on seed germination & seedling growth of Rabdosia rubescens].

The seeds of Rabdosia rubescens were as the materials to research the impacts of different lead (Pb2+) concentrations(0, 135, 270, 540, 1 080 mg x L(-1)) on seed germination and seedling growth. The results show that: Low concentration of lead had no obvious effect on early germination of the seed, the germination vigor and germination speed were lightly higher but not significantly differed at the level of Pb concentration 135 mg x L(-1) with control group; Mid-high concentration of Pb solution (270-1 080 mg x L(-1)) significantly inhibited the seed germination and seedling growth, which reduced the seed germination rate, germination vigor, germination index, embryo root length and shoot length, growth index with increasing of Pb concentrations. There was a inhibitory effect on embryo shoot length and root length at mid-high lead concentrations stress, and stronger inhibitory effect on root , which was more sensitive than shoot to Pb stress(P < 0.05). Pb bioaccumulation coefficient (BC) was 0.76-2.59, increased with concentration of Pb; Pb enrichment in seedling mainly caused the growth inhibition. The fitting model predictive analyses show, the critical concentration of Pb, which causes the germination rate and biomass fresh weight reducing 10%, is 195.18, 101.65 mg x L(-1).

[Determination of mineral elements in cistanche and its extractives by ICP-AES].

Determination of Fe, Zn, Cu, K, P, Ca, Mg, Na, B and Al in cistanche and its extractives was carried out by the inductively coupled plasma atomic emission spectrometry (ICP-AES). The results showed that (1) the contents of Mn, K, Ca, Mg and Na in cistanche deserticola Y. C. Ma were 17.70, 16,278. 03, 1,947.25, 1,662.76, and 106.79 respectively (microg.g-1), much higher than in Cistanche tubulosa (Schrenk) Wight; (2) the contents of Fe, Cu and P in 50% methanol extract of Cistanche deserticola were 1.5, 2.4, and 1.1 times that of aqueous extract respectively, the contents of Mn, Zn, K, P, Mg and Na were 3.8, 1.2, 1.6, 2.3, 2.2, and 1.1 times that of methanol extract respectively; (3) the contents of Cu and P in 50% methanol extract of Cistanche tubulosa were 1.6 and 1.2 times that of aqueous extract, the contents of Mn, Zn, K, P, Mg, Na and Al were 2.0, 1.6, 1.6, 1.4, 1.8, 1.2, and 1.2 times that of methanol extract respectively; (4) the leaching rate of 11 mineral elements in Cistanche deserticola and Cistanche tubulosa were 60.79%-83. 10% and 44. 58%-83.84% in 50% methanol respectively, and the leaching rate of Mn, Zn, K, P, Ca, Na and Al were much higher than in water and 100% methanol extracted. The research provided the basic data of mineral element in comparative study on Cistanche deserticola and Cistanche tubulosa, and provide scientific basis for efficient utilization of mineral element in cistanche.

[Simultaneous determination of seven compounds in Lonicera japonica by high performance liquid chromatography].

A high performance liquid chromatographic (HPLC) method was developed to determine the contents of seven compounds in Lonicera japonica and Folium Lonicerae with maximum wavelength conversion program, and analyze the content differences between them. The separation was performed on an Agilent Eclipse Plus C18 column (250 mm x 4.6 mm, 5 microm) operated at normal temperature with the gradient elution by two mobile phases of water (including 0.3% (v/v) formic acid) (A) and acetonitrile (B) at a flow rate of 1 mL/min. The maximum detection wavelength was set at 330 nm and 350 nm by conversion. The contents of chlorogenic acid and caffeic acid in Lonicera new leaves were 2.572% and 1.498 per thousand respectively, both higher than those cited in Chinese Pharmacopoeia, indicating that Lonicera new leaves have the necessity to be further studied and developed. This method is simple, rapid and highly sensitive. It is suitable for the simultaneous determination of the seven compounds and the quality control of Lonicera japonica.

[Analysis of mineral elements in different organs of Chrysanthemum indicum L. based on ICP-AES].

ICP-AES technique was used to analyse the mineral elements in different organs of Chrysanthemum indicum L. The results show that: (1) The content of K in root, stem, leaves and flower was the highest among macroelements, and can respectively reach to 15.84, 17.74, 31.52 and 37.55 mg x g(-1), while the content of Fe was the highest in microelements in root, stem, leaves and flower, and can respectively reach to 3 219.90, 433.36, 1 519.46, and 1 426.63 microg x g(-1). (2) The accumulation of K, P, Ca, Mg, Mn, Zn and Mo was highest in stem of Chrysanthemum indicum L, and can respectively reach to 240.61, 19.67, 74.87, 18.31 mg x plant(-1) and 893.00, 1 039.08, 2.85 microg x plant(-1), while the accumulation of Na, Fe and Cu was highest in root, and can respectively reach to 11.51 mg x plant(-1), 11 725.27 microg x plant(-1), and 235.24 microg x plant(-1). (3) The proportion of K:P, Ca:Mg, Fe:Mn, Zn:Cu was varied in different organs. The results of the study pinpoint the mineral elements in different organs of Chrysanthemum indicum L. and will be a scientific basis for fertilization.

Echinacoside Stimulates Cell Proliferation and Prevents Cell Apoptosis in Intestinal Epithelial MODE-K Cells by Up-Regulation of Transforming Growth Factor-ß1 Expression.

Cistanche deserticola MA (C. deserticola) has been widely used as a laxative herbal in herbal medicine for the treatment of irritable bowel syndrome or constipation, and echinacoside (ECH) is one of the major bioactive ingredients in this herbal. Our aim was to investigate the effect of ECH on intestinal epithelial cell growth and death. MODE-K, an intestinal epithelial cell line, was used as an in vitro model of the intestine. Cell proliferation was measured by methylthiazol tetrazolium (MTT) assay. Cell apoptosis was determined with Annexin-V staining. Here we showed that in cultured MODE-K cells, ECH significantly stimulated cell proliferation and enhanced cell survival by reducing cell apoptosis in the presence of H2O2 or the mixture of pro-inflammatory cytokines, while transforming growth factor (TGF)-ß1 expression was up-regulated in a dose-dependent manner. Knockdown of TGF-ß1 expression disrupted both the proliferative and cytoprotective activities of ECH, which was further confirmed by neutralization of TGF-ß1 activity using anti-TGF-ß1 antibody. These data suggest that ECH as one of bioactive ingredients in herbal C. deserticola and others may improve mucosal tissue repair by stimulating intestinal epithelial cell proliferation and preventing cell death via up-regulation of TGF-ß.

Echinacoside stimulates cell proliferation and prevents cell apoptosis in intestinal epithelial MODE-K cells by up-regulation of transforming growth factor-ß1 expression.

Cistanche deserticola MA (C. deserticola) has been widely used as a laxative herbal in herbal medicine for the treatment of irritable bowel syndrome or constipation, and echinacoside (ECH) is one of the major bioactive ingredients in this herbal. Our aim was to investigate the effect of ECH on intestinal epithelial cell growth and death. MODE-K, an intestinal epithelial cell line, was used as an in vitro model of the intestine. Cell proliferation was measured by methylthiazol tetrazolium (MTT) assay. Cell apoptosis was determined with Annexin-V staining. Here we showed that in cultured MODE-K cells, ECH significantly stimulated cell proliferation and enhanced cell survival by reducing cell apoptosis in the presence of H(2)O(2) or the mixture of pro-inflammatory cytokines, while transforming growth factor (TGF)-ß1 expression was up-regulated in a dose-dependent manner. Knockdown of TGF-ß1 expression disrupted both the proliferative and cytoprotective activities of ECH, which was further confirmed by neutralization of TGF-ß1 activity using anti-TGF-ß1 antibody. These data suggest that ECH as one of bioactive ingredients in herbal C. deserticola and others may improve mucosal tissue repair by stimulating intestinal epithelial cell proliferation and preventing cell death via up-regulation of TGF-ß.

Reduction of inflammatory hyperplasia in the intestine in colon cancer-prone mice by water-extract of Cistanche deserticola.

Cistanche deserticola has commonly been used in traditional Chinese medicine to treat many health problems including irritable bowel syndrome or constipation. This study was designed to test the efficacy of a water-extract of C. deserticola in the prevention of colorectal cancer in a mouse model. Polysaccharide-rich water-extract of C. deserticola was prepared by boiling its stem powder in distilled water. Tgfb1Rag2 null mice were used as an experimental model. Here we showed that feeding of water-extract of C. deserticola significantly reduced the number of mucosal hyperplasia and intestinal helicobacter infection in mice. This beneficial effect correlated with significant stimulation of the immune system, evidenced by the enlargement of the spleens with increased number of splenic macrophage and natural killer cells, and with more potent cytotoxicity of splenocytes. In vitro water-extract of C. deserticola enhanced the cytotoxicity of naïve splenocytes against a human colon cancer cell line, and in macrophage cultures up-regulated nitric oxide synthase II expression and stimulated phagocytosis. In conclusion, our data indicate that oral administration of C. deserticola extract reduces inflammatory hyperplastic polyps and helicobacter infection in mice by its immune-stimulatory activity, suggesting that C. deserticola extract may have potential in preventing intestinal inflammation disorders including colorectal cancer.

[Analysis of the content of mineral elements in Scutellaria baicalensis, skullcap tea and its solution].

The content of nine mineral elements in the root, stem and leaf of 1-3 years old Scutellaria baicalensis, skullcap tea, and its solution was determined by the inductively coupled plasma atomic emission spectrometry (ICP-AES). The results show that the main mineral elements in the root, stem and leaf are similar. The main mineral elements include K, Ca, Mg, P, Al and Fe. The content ratio of Zn, Mn, Cu, Fe, Al, P, Mg, Ca and K in the leaf of Scutellaria haicalensis is 1 : 3 : 6 : 15 : 18 19 : 41 : 333 : 423. The contents of Fe, Mn, P, Mg and Ca in the stems and leaves of Scutellaria baicalensis increase with the increase of cultivation period. There were K, Ca, Mg and other mineral elements in the skullcap tea, and Fe, Zn and Mg were easy to be dissolved, the dissolution rates were 61.8%, 55.4% and 61.4% respectively. This study can provide scientific basis for rational utilization of the above ground resources of Scutellaria baicalensis.

[Determination of trace elements in holoparasitism Cistanche tubulosa].

The trace elements in Chinese herbal medicines are important nutritional elements for human health. Application of ICP-AES was applied to detect Fe, Mn, Cu, Zn and Mg of the different parts and size grading on growth of Cistanche tubulosa. The results showed that there were comparatively rich trace elements in Cistanche tubulosa, and the trace elements of the different parts and size grading on growth were significantly different. Traditionally, the succulent stem and 200-500 grams of Cistanche tubulosa are common in the use of medicinal herbs, and the authors found that the iron, manganese, copper, zinc and magnesium were 242.18, 17.85, 7.96, 6.64 and 1 357.50 microg x g(-1) in the succulent stem of Cistanche tubulosa. There were 174.21, 4.48, 8.11, 4.31 and 1 126.93 microg x g(-1) in 200-500 grams of Cistanche tubulosa. It provided useful data for discussing the relationship between the content of the trace elements and the efficacy of Cistanche tubulosa.

[Analysis on Cistanche tubulosa that parasites on different Tamarixs by RAPD].

OBJECTIVE: To study genetic difference of Cistanche tubulosa that parasites on different Tamarixs and give a reference to select host of C. tubulosa. METHOD: Sixteen selected primers by random amplified polymorphic DNA (RAPD) markers were used to analyze genetic distance of C. tubulosa that parasites on eight different hosts. RESULT: Sixty-six point seven percent of the total bands were polymorphic, that proved the genetic diversity level in different C. tubulosa types was relatively high, especially the two that parasites on Tamarix hispida and T. chinensis. Cultural areas had more remarkable influence on genetic distance of Cistanche tubulosa than the hosts, and introduction was helpful to maintain the more genetic diversity in different C. tubulosa types. Genetic difference in different C. tubulosa types was far less than that between different species in Cistanche. CONCLUSION: C. tubulosa types which parasite on different Tamarixs have high genetic diversity.

[Study on conditions of seed germination of Cistanche].

OBJECTIVE: To study the effect of fluridone concentration, stimulating period, temperature and salt on the seed germination of three species of Cistanche. METHOD: The seeds were cultured in Petri dish, and the germination percentage was counted. RESULT: The highest germination percentage was observed in Cistanche tubulosa, C. deserticola, C. sala seeds pre-treated by 0.1 mg x L(-1) fluridone for 24-29 h. The optimal temperature for the seeds germination of three species of Cistanche was at 20-30 degrees C, and the seeds did not germinate at sub-or supraoptimal temperatures (5 and 35 degrees C). The salt tolerance of C. sala seeds was strong, and the critical value of NaCl concentration was 0.04 mol x L(-1). By contrast, C. tubulosa and C. deserticola seeds were more sensitive to the salt stress, the critical value of NaCl concentration was 0.02 mol x L(-1). CONCLUSION: The optimal germination condition and the method of testing germination percentage of three species of Cistanche seeds are as follow: the seeds are pre-treated by 0.1 mg x L(-1) fluridone for 24 h and then cultured at 20-30 degrees C in salt solution which concentration is lower than 0.02 mol x L(-1).

[Studies on accumulation of active ingredients, N, P and K in Anemarrhena asphodeloides].

OBJECTIVE: To study the accumulation of active ingredients, the absorption and transformation of N, P and K in Anemarrhena asphodeloides and provide basis for determination of the harvest time and fertilizing. METHOD: Samples were collected in different phrases and the weight of dry matter, the content of N, P and K of different organs and the content of sarsasapogenin were determined. RESULT: Absorption of N, P and K started by the root and rhizoma after July. At the end of August, the N and K of the aerial part transfered largely into rhizome. The content of sarsasapogenin in rhizome was the highest in early spring. CONCLUSION: Additional fertilizer is helpful to increase the yield in July of the second year after the transplantation. The quality is the best when harvest in early spring.

[Studied of dry matter accumulation and echinacoside content of Cistanche tubulosa in Huabei plain].

OBJECTIVE: To give some theory support of Cistanche tubulosa cultivation by searching dry matter accumulation and echinacoside content of C. tubulosa. METHOD: Dry matter accumulation content of C. tubulosa culturing in Huabei plain was analysed in different growth season of C. tubulosa. Echinacoside content was determined by HPLC. RESULT: Dry matter accumulation of C. tubulosa showed "S" variation. Dry matter accumulation increased fastest in September among growing seasons. Dry matter amount was 138.58 g after C. tubulosa grew a year. Dry matter amount decreased significantly along with inoculation time retarded. Echinacoside content was 30.59% when C. tubulosa grew in 5 months, decreased guadully after that, and 9.76% in annual. CONCLUSION: Variation rule of dry matter accumulation and echinacoside content was found in C. tubulosa that grew one year in Huabei plain.

[Studies on cultivation measures of Angelica dahurica].

OBJECTIVE: To evaluate the effect of seedingtime, density of crop and fertilization on the yield of Angelica dahurica. METHOD: Use weighing method to measure the output of A. dahurica. RESULT: The highest yield of seeding-time is 8373 kg/hm' on April 20, which is considerably different compared with April 5 and May 5; the highest yield of the density is 9300 kg/hm2 on 330,000 plants/hm2; the yield of fertilization tests all are considerable higher than that of the contrast. CONCLUSION: The appropriate seeingtime of A. dahurica is the first or second ten days of April, the appropriate density is 330,000 plants/hm2, and the appropriate amount of fertilization is N24P20, i.e pure N 360 kg and P20, 300 kg per hectare.

[Effects of cultivation technique on yield and favonoid content of Chrysanthemum flower (Qiju) grown in Hebei].

OBJECTIVE: To study the effect of cultivation techniques on the flower yield flavonoid content in Chrysanthemum flower grown in Hebei. METHOD: Studied on flowers yield and three factors (transplanting date and plant density and fertilizer quantity) were examined in field experiment at 4 treatments levels. RESULT AND CONCLUSION: The best results were obtained at following conditions: diammonium phosphate 300 kg x hm(-2) and potassium sulfate 150 kg x hm(-2) fertilized before transplanting, transplanting at the first ten days of May and the spacing 40 cm x 40 cm.

[Study on inoculation technology of Cistanche tubulosa in the field].

OBJECTIVE: To increase inoculation rate of Cistanche tubulosa in the field by studying inoculation technologies. METHOD: Root-tube inoculation methed was used on field experiments. Inoculation rate of C. tubulosa was compared to different size seeds and inoculation mediums and inoculation time. RESULT AND CONCLUSION: May is suitable inoculation time. The inoculation rate of C. tubulosa is 92.5% while the seed width is more than 0.7 mm and coarse sand is selected during inoculation period.

[Morphology of seed germination and parasitism in Cistanche tubulosa].

OBJECTIVE: To understand the process of Cistanche tubulosa. METHOD: The process of seed germination and parasitism was observed using stereomicroscope. RESULT: Seedling of C. tubulosa sprouted after forty day without host root's contact in fields, a tube-like-organ formed and grew auger-type from host root, the tuber apex where touches host root swelled and formed haustorium. Haustorium intruded host root epidermis and vascular bundles, and released brown substances. Then, embryo bud with six or more young leaves formed, finally the swelled tuber-like-organ broken and seed coat shed. Due to the parasitism of C. tubulosa, the host root near stem site swelled, but the other part, shrunk and disappered gradually. CONCLUSION: Seed of C. tubulosa could germinate indepently in fields. Tuber-like-organ formatin, haustorium formation and bud formation are key steps of C. tubulosa seedling development.