OBJECTIVETo study the effects of methyl jasmonate (MJ) on the accumulation and release of tanshinones in suspension cultures of Salvia miltiorrhiza hairy roots.

METHODAfter 18 day's suspension culture of S. miltiorrhiza hairy roots induced by Agrobacterium rhizogenes ATCC15834, the chemical elicitor--methyl jasmonat was added into 6-7V suspension cultures and at the same time, tanshinones contents (including cryptotanshinone and tanshinone II(A)) on the day 2, 6 and 9, after dealing with MJ, was quantified by HPLC.

RESULTAfter dealing with MJ on the day 2, 6 and 9, the concentration of cryptotanshinone reached to 0.039, 0.204, 0.571 mg x g(-1) respectively,and tanshinone II(A) reached 0.251, 0.601 and 1.563 mg x g(-1) respectively. After 9 day's treatment by MJ, the maximum increase of cryptotanshinone and tanshinone II(A) were 23.8 fold and 6.2 fold higher than that of the control respectively.

CONCLUSIONMJ could stimulate the accumulation of tanshinones in S. miltiorrhiza and have released them into the culture medium.

Acetates ; pharmacology ; Culture Techniques ; Cyclopentanes ; pharmacology ; Diterpenes, Abietane ; Oxylipins ; pharmacology ; Phenanthrenes ; metabolism ; Plant Growth Regulators ; pharmacology ; Plant Roots ; drug effects ; growth & development ; metabolism ; Plants, Medicinal ; drug effects ; growth & development ; metabolism ; Salvia miltiorrhiza ; drug effects ; growth & development ; metabolism

A new phenylpropanoid (1), together with seven known ones (2-8), has been isolated from the flowers of Rosa rugosa collected from Shanxi province by using various chromatographic techniques. Compound 1 is a new compound, and it displayed cytotoxicity against NB4, SH-SY5Y, PC3, A549 and MCF7 cell lines with IC₅₀ values of 8.2, 6.2, 4.3, 2.8, and 9.6 µmol · L⁻¹ respectively.
Cell Line ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Flowers ; chemistry ; Humans ; Molecular Structure ; Phenylethyl Alcohol ; chemistry ; isolation & purification ; pharmacology ; Rosa ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVEStudying the gene expression profiling of different stage hairy root of Salvia miltiorrhiza, in order to find functional genes.

METHODThe contents of second metabolites were determined by HPLC and gene expression profiling was detected by cDNA microarray. cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine's linear RNA amplification method and subsequent enzymatic reaction. The microarrays were scanned with a ScanArray Express scanner using ScanArray 2.0 software and quantified by signal intensities of individual spots from the 16-bit TIFF images using GenePix Pro 4.0. The linear normalization method was used for data analyze. Northern blot was used to test the gene expression results obtained by microarray. Different expressed genes were sequenced and analyzed by gap4 software, and then they were analyzed with BLASTX, BLASTN, GO and KEGG.

RESULTGrowth rate and second metabolites analysis indicated that the stage from 30 d to 45 d was the growth stage, while the stage from 45 d to 60 d was the second metabolites accumulation stage. Accordingly 30 d hairy root was chosen as a reference, which was hybridized with 45 d and 60 d hairy root separately. Total 203 different expressed genes were obtained. Northern blot showed that the result was identical with the microarray result. After sequenced, there were 172 genes clustered into 114 clusters (Unigenes). Among them, 62 unigenes had known functions, 34 unigenes were hypothetical protein, 9 unigenes were homologues with no similarity and 9 unigenes were unidentified protein with low similarity. Total 67 genes were classified into cellular component ontology, molecular function ontology and biological process ontology based on GO analysis. Total 26 genes, which represented 29 metabolic-related enzymes, were located in metabolic maps based on KEGG pathway classification.

CONCLUSIONSeveral important functional genes related to second metabolite synthesis were cloned such as P450 and copalyl diphosphate synthase genes. cDNA microarray was a useful tool for functional genomics of traditional Chinese medicine.

Alkyl and Aryl Transferases ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Salvia miltiorrhiza ; genetics ; growth & development ; metabolism

OBJECTIVETo study on the chemical stability of Salvia miltirrhiza hairy root.

METHODThe rolA gene was detected by PCR in DNA and the chemical contituent variances were detected by HPLC.

RESULTThe rolA gene was found in all the 10 batches of the culfured hairy root. The similarities of the chromatographic fingerprints of the 10 batches are higher than 0. 95.

CONCLUSIONThere are no significant differences of the chemical constituents in 10 hairy root samples.

Chromatography, High Pressure Liquid ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; Genes, Plant ; Plant Roots ; chemistry ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; genetics ; Sensitivity and Specificity

OBJECTIVETo investigate the role of mesenchymal stem cells (BMSCs) in multiple myeloma (MM) bone marrow (BM) microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.

METHODSBMSCs were derived from BM of untreated myeloma patients (MM-BMSCs) and healthy donors (HD-BMSCs), respectively. The phenotype, proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs. Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro. The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.

RESULTSMM-BMSCs and HD-BMSCs were isolated successfully. The phenotype of MM-BMSCs was similar to that of HD-BMSCs. Expressions of CD73, CD105, CD44 and CD29 were positive, but those of CD31, CD34, CD45 and HLA-DR (< 1%) negative. The proliferation time of MM-BMSCs was longer than that of HD-BMSCs (82 h vs 62 h, P < 0.05). Moreover, over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ± 9.4) pg/ml vs (115.0 ± 15.1) pg/ml and (1497.2 ± 39.7) pg/ml vs (1329.0 ± 21.1) pg/ml, respectively]. MM- BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.

CONCLUSIONSMM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib. Over-expression of IL-6 and VEGF maybe play a critical role in these effects.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Bortezomib ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Multiple Myeloma ; metabolism

OBJECTIVEEstablishing cDNA microarray, in order to study functional genomics of Salvia miltiorrhiza.

METHODTotal RNA samples were prepared from S. miltiorrhiza roots using a modified CTAB method. mRNA was isolated by Quichprep Micro mRNA Purification Kit from Pharmacia. Then cDNA was synthesized and cloned into the EcoRI-XhoI sites of the ZAP Express vector using a cDNA synthesis kit, and the ligation mixture was packaged using a ZAP-cDNA Gigapack Gold III cloning kit (Stratagene). The single phage was isolated for PCR amplification, Aliquots of the PCR reactions were analyzed in a 1.5% agarose gel to verify the quality of PCR. The remaining cDNA was purified by Multiscreen filter plates (Millipore) and aliquots were analyzed by agarose gel again to verify the quality of purification. Clones passed verification was resuspended in 15 microL 50% DMSO for arraying. An actin gene from S. miltiorrhiza was used for positive control. PloyA and 50% DMSO was used for negative controls.

RESULTBacterial colonies containing cNDAs of S. miltiorrhiza were inserted with average insert size of 0. 5 kb to 2. 5 kb. Total 4 354 genes were singled out from the first 8 736 PCR product and used for cDNA microarray manufacture. Single color fluorescence hybridization showed that all positive controls had signals while negative controls had no signals.

CONCLUSIONIt was the first cDNA microarray about traditional Chinese herbs especially for geoherbs. It could be a powerful tool for studying functional genomics of S. miltiorrhiza.

Gene Expression Profiling ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; methods ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Salvia miltiorrhiza ; genetics

OBJECTIVESTo investigate the possibilities of an association between the degrees of HBV suppression with nucleoside treatments at week 24 and week 52 in hepatitis B patients and to find a useful predictor for treatment efficacy.

METHODSIn this phase III, double-blind, multicenter trial, we compared the efficacy of telbivudine treatment with lamivudine treatment in 332 Chinese compensated chronic hepatitis B patients. The patients were randomly assigned to a daily 600 mg telbivudine treatment group or daily 100 mg lamivudine group for 24 weeks. They were then categorized into 4 groups according to their serum HBV DNA levels (copies/ml) at week 24: a PCR-undetectable group (< 300 copies/ml); a QL- < 10(3) copies/ml group; a 10(3)-<10(4) copies/ml group; and a > or = 10(4) copies/ml group. The treatments were continued as they previously had been for another 28 weeks and the patients serum HBV DNA levels were examined again.

RESULTSAt week 52, mean reductions of serum HBV DNA were significantly greater in the telbivudine-treated patients than in the lamivudine-treated group (6.2 log10 vs 5.4 log10, t = 3.6, P < 0.01). Viral resistance was twice as common in lamivudine-treated patients compared to those receiving telbivudine. Telbivudine was well-tolerated with an adverse event profile similar to that of lamivudine. The lower the HBV DNA level achieved at week 24, the higher HBV DNA non-detectable by PCR. ALT normalization and HBeAg seroconversion achieved at week 52, and viral resistance at week 48 decreased parallel to the degree of HBV DNA inhibition.

CONCLUSIONHBV DNA PCR-undetectable at week 24 in nucleoside-treated hepatitis B patients suggests a better efficacy at week 52 and lower viral resistance at week 48. The degree of suppression of HBV at week 24 may be used as a predictor of 1-year outcome.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; Double-Blind Method ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Male ; Middle Aged ; Nucleosides ; therapeutic use ; Pyrimidinones ; therapeutic use ; Thymidine ; analogs & derivatives ; Treatment Outcome ; Young Adult

Background: Rapid visual acuity (VA) decline was a common complaint in patients with sellar/suprasellar germinoma. In our hospital, 3.4 Gy/2f of emergency irradiation was applied to save patient VA and enable subsequent chemoradiotherapy. This study aimed to investigate the efficacy of emergency irradiation with 3.4 Gy/2f in patients with sellar/suprasellar germinoma who had rapid VA decline. Methods: From January 2014 to December 2017, 33 patients with sellar/suprasellar germinoma who complained of VA decline within 3 months received 3.4 Gy/2f of emergency irradiation in Beijing Tiantan Hospital. The best-corrected VA (BCVA) and mean deviation (MD) were measured. Correlations between visual function change and clinical factors, including age at diagnosis, duration of VA decline, extent of tumor regression, serum level of tumor markers, were analyzed. Results: Among 33 patients with sellar/suprasellar germinoma, the median diameter and volume of sellar/suprasellar lesions were 32 mm (range: 5–55 mm) and 12.9 cm³ (range 0.6–58.5 cm³), respectively. Data on pre- and post-emergency-irradiation BCVA were obtained in 32 patients. For the right eyes, BCVA was improved in 23 patients (71.9%), unchanged in 7 (21.9%), and worsened in 2 (6.2%); and for the left eyes, these numbers were 27 (84.4%), 4 (12.5%), and 1 (3.1%), respectively. In terms of the logarithm of the minimum angle of resolution (logarithm of the minimum angle of resolution = Log (1/BCVA) score, the improvement was significant in both eyes (P < 0.001). In terms of MD, six patients had paired data and the improvement was marginal in the right eyes (P = 0.068) and significant in the left eyes (P = 0.043). However, no clinical factor was found to have correlation with visual function improvement. Conclusion In sellar/suprasellar germinoma patients with VA decline, 3.4 Gy/2f of emergency irradiation was effective in improving visual function.

Objective:To review the categories of physical activities and rehabilitation exercise for aging people based on the theory and method of the International Classification of Functioning, Disability, and Health (ICF). Methods:From the framework and coding of ICF, physical activities, rehabilitation exercise and functional improvement for the aging people were reviewed. Results:There were three typical physical activities: physical fitness, skills and sports, mainly including regular exercises, cognitive amusements, leisure sports activities, recreational sports activities, rehabilitation exercises, etc. Physical activities promoted the functioning of the aging people, including mental function (b1), pain and sensory dysfunction (b2), cardiovascular, blood, the function of the immune system and respiratory system (b4), digestion, metabolism and function of the endocrine system (b5), nerve musculoskeletal function related to movement (b7), the function of skin and related structures (b8), activity (d4), and community, social and civic life (d9), etc., especially body mass index, cardiovascular, physical fitness (muscular strength and endurance, aerobic endurance), balance, flexibility, upper and lower extremities strength, sleep, metabolic capability, cognitive function and anti-aging ability. There were more gains at individual levels, including promotion of subjective happiness, quality of life, reducing depression, risk of sarcopenia, dementia and falling, etc. The environmental and personal factors related to activity and participation in physical activities included the products and technologies (e1), natural environment and man-made changes to the environment (e3), support and interpersonal relationships (e3), attitude, service system and policy (e5), such as urban environment, building environment, street pavement behavior, weather, caregivers, accompany of family and friends, etc. Conclusion:The physical activities and rehabilitation exercise the aged joined include physical fitness activities, skills activities, and sports activities. The physical activities had effects on the rehabilitation of the aged including the function of the body(mental function, pain and sensory dysfunction, cardiovascular, blood, the function of the immune system and respiratory system, digestion, metabolism, and function of the endocrine system, nerve musculoskeletal function related to movement, the function of skin and related structures) and the participation in activities (activities, community, society and civic life). The function of the body works for the health of the aged to promote physical health, mental health, functional health, social adjustment and social well-being.