A full-length cDNA encoding a MYB-related regulatory gene was isolated from a cDNA library prepared from mRNAs of the red line callus of S. medusa by TD-PCR. The cDNA, designated SmP, is 969 nucleotides long and has an open reading frame of 771 bp with a deduced amino acid sequence of 256 residues. The putative protein of SmP has two typical conversed R2R3-Myb DNA-binding domains in N-terminal and displays a rather high degree of similarity to OsMYB from rice and LBMI from tobacco, showing 73% and 70% identity within the DNA-binding domains. However, the C-terminal domain of the SmP protein does not show obvious similarity to any other known protein sequence. It is rich in hydrophilic amino acids, especially in serine residues (18.38%), partly organized in homopolymeric stretches, a feature often found in activation domain of transcription factors.
Amino Acid Sequence ; Cloning, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; classification ; genetics ; metabolism ; Polymerase Chain Reaction ; Saussurea ; classification ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Transcription Factors ; chemistry ; genetics ; metabolism

OBJECTIVETo establish an HPLC fingerprint of Rhizoma fagopyri dibotoryis.

METHODThe HPLC-electrochemical detection assay was used to establish the fingerprint of Rhizoma fagopyri dibotoryis. The sample was performed on a column of Diamonsil C18 (4.6 mm x 250 mm, 5 microm) which was eluted with methanol- 0.1 mol x L(-1) phosphate buffer (pH 2.5), the flow rate was 1.0 mL x min(-1), the column temperature was 35 degrees C, the reference electrode was ISAAC (in-situ silver/silver chloride), the work electrode was glassy carbon, the counter electrode was Pt platmun.

RESULTThe HPLC fingerprint profiles of 6 Rhizoma fagopyri dibotoryis contains 6 common chromatographic peaks, and gallic acid, protocatechuic acid, protocatechuic aldehyde and ( - ) -epicatechin were tested the samples. The contents of protocatechuic acid and protocatechuic aldehyde were from 0.004% to 0.05% and from 0.003% to 0.015%, respectively.

CONCLUSIONThe method can be used to control the quality of Rhizoma fagopyri dibotoryis.

Benzaldehydes ; analysis ; Catechin ; analysis ; Catechols ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; standards ; Electrochemistry ; Fagopyrum ; chemistry ; Gallic Acid ; analysis ; Hydroxybenzoates ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Rhizome ; chemistry

OBJECTIVEAIM To establish a drug screening model based on transcriptional regulation of estrogen responsive element (ERE) and use it to screen compounds for discovering new ligands of estrogen receptor (ER) subtypes.

METHODA recombinant reporter vector pERE-TAL-SEAP was constructed by inserting a synthetic sequence composed of five tandem copies of EREs upstream of promoter of the reporter vector pTAL-SEAP. The pERE-TAL-SEAP and the internal control plasmid pCMV were transiently co-transfected into Hela cells expressing ER subtype or ER subtype, and the effects of pure ER agonists 17estradiol, phytoestrogen genistein and pure ER antagonist ICI182, 780 on reporter gene SEAP expression were observed.

RESULTIn the Hela cells expressing ER alpha or ER beta subtype, the expression of SEAP gene were induced in a dose dependent manner by 17-estrodiol with a maximal effect at approximately 10 nmol.L-1 and with EC50 of (80.58 +/- 8.51) pmol.L-1 and (103.90 +/- 5.29) pmol.L-1, respectively, so done by phytoestrogen genistein with a maximal effect at 1 mumol.L-1 and with EC50 of (10.86 +/- 0.75) nmol.L-1 and (39.38 +/- 2.26) nmol.L-1, respectively. The maximal level induced by estrodiol and genistein were about 7-14 fold higher than that of vehicle. The pure antiestrogen ICI182, 780 at concentration of 1 mumol.L-1 completely blocked the inductions of 17-estrodiol and genistein.

CONCLUSIONThe cellular drug screening model can be established by transfecting reporter vector pERE-TAL-SEAP in Hela cell lines expressing ER alpha or ER beta. The cell lines can be used to screen compounds with estrogenicity by testing SEAP activity in the culture media of cells growing in microtitier wells. The system should provide an efficient model for screening and analyzing the activity of large numbers of ligands of ER.

Drug Evaluation, Preclinical ; methods ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; Estrogen Receptor beta ; Gene Expression Regulation ; drug effects ; Genes, Reporter ; Genistein ; pharmacology ; HeLa Cells ; Humans ; Ligands ; Promoter Regions, Genetic ; Receptors, Estrogen ; genetics ; Transfection

OBJECTIVETo establish a HPLC fingerprint for quantitative analysis of the active constituents of jizhi syrup.

METHODHPLC analysis was performed on a Zorbax Sb C18 column (4.6 mm x 250 mm, 5 microm), the mobile phase in gradient elution was composed of (A) 1% acetic acid solution (including 0.2% triethylamine) and (B) acetonitrile, at a flow rate of 1.0 mL x min(-1). The column temperature was 30 degrees C and the wavelength was 280 nm.

RESULT21 common peaks were tested in 10 batches of samples. Compared with the standard fingerprint, the similarity of each sample was greater than 0.99. At the same time, protocatechuic acid, protocatechu aldehyde, chlorogenic acid, caffeic acid, naringin and neohesperidin were found in all samples.

CONCLUSIONThe established method can be used for the quality control of jizhi syrup.

Antitussive Agents ; chemistry ; Benzaldehydes ; analysis ; Catechols ; analysis ; Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavanones ; analysis ; Hydroxybenzoates ; analysis ; Plants, Medicinal ; chemistry ; Reproducibility of Results

Objective:To analyze the effects of two decomposition algorithms of dual-energy cone beam CT (DECBCT) (direct decomposition and iterative decomposition) on the image quality and material decomposition accuracy of different sizes of phantoms.Methods:Different sizes of imaging parts of patients were simulated using the combination of CatPhan604 phantoms and customized annuluses. CBCT with high energy of 140 kVp and low energy of 100 kVp were acquired using the Varian Edge CBCT system. Then the material decomposition of DECBCT images was performed using the two algorithms. The electron density (ED) and contrast-to-noise ratio (CNR) of each material in the CTP682 module were calculated. They were used to assess the decomposition accuracy and image quality of the two algorithms.Results:Based on the values in the Catphan604 manual, both algorithms have high ED accuracy. Only the ED accuracy of four materials of the smallest sized phantom showed statistical difference ( z = -4.21, 4.30, 2.87, 5.45, P < 0.05), but the average relative error was less than 1%. The CNR of the iterative decomposition algorithm was significantly higher than that of the direct decomposition, increasing by 51.8%-703.47%. The increase in the phantom size significantly reduced the accuracy of ED, and the increased amplitude of the relative error was up to a maximum of 2.52%. The large phantom size also reduced the image quality of iterative decomposition, and the decreased amplitude of CNR was up to a maximum of 39.71. Conclusions:Compared with the direct decomposition, the iterative decomposition algorithm can significantly reduce the image noise and improve the contrast without losing the accuracy of electron density in the DECBCT construction of different sizes of phantoms.

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Acupuncture Points ; Acupuncture Therapy ; Human Body ; Humans ; Meridians

As a traditional Chinese medicinal material， Glycyrrhizae Radix et Rhizoma has been extensively used in various formulae. According to modern pharmacological research， it has anti-tumor， anti-inflammatory， anti-viral， liver-protecting， anti-heart failure， immunoregulatory， and anti-fibrosis effects. Caused by the interaction of various factors， cancer features complex pathogenesis. It is a global challenge and one of the main causes of death in China. Statistics show that the morbidity and mortality of malignant tumors have been on the rise， particularly for the young， which threaten the health of human beings. At the moment， radiotherapy and chemotherapy are the main countermeasures. Most clinical anti-tumor drugs demonstrate non-selective toxicity. To be specific， they damage normal cells while killing tumor cells， thus injuring vital organs. In addition， long-term medication will reduce the sensitivity of tumor cells. However， traditional Chinese medicine， which is characterized by treatment based on syndrome differentiation， multiple components， and multiple targets， is superior in the treatment of tumor. Studies have shown that the combination of anti-tumor drugs with Chinese medicine can not only enhance the anti-tumor effect but also alleviate toxicity. Therefore， it has been a research hotspot to develop anti-tumor drugs based on traditional Chinese medicine. In recent years， major headway has been made in the research on active ingreddients of Glycyrrhizae Radix et Rhizoma in anti-liver cancer， anti-breast cancer， anti-lung cancer， and anti-colon cancer and the combination with other drugs for anti-tumor. On this basis， we summarized the mechanisms of active ingredients of Glycyrrhizae Radix et Rhizoma in inducing apoptosis， interfering with cell cycle， inducing autophagy， inhibiting glycolysis， regulating immunity， and modulating miRNA and signaling pathways， as well as the combination with other drugs in anti-tumor efficiency， toxicity reduction， and sensitivity enhancement， hoping to lay a theoretical basis for the further development and clinical application of active ingredients of Glycyrrhizae Radix et Rhizoma.

Pathological diagnosis of salivary gland tumors is one of the most challenging areas in all head and neck surgical pathology. The classification of salivary gland tumors was updated in the 5th edition of the World Health Organization Classification of Head and Neck Tumours, most of which were based on their molecular pathological characteristerics. This new classification features a description of several new entitiesamong benign and malignant neoplasms, salivary gland tumors with updated naming or diagnostic criteria, and lesions deleted from this section, etc.This present review focuses on the updates and changes in the new classification of salivary gland tumors, and provides some reference for head and neck surgeons and pathologists.
Humans ; Head and Neck Neoplasms ; Salivary Gland Neoplasms/pathology* ; Salivary Glands ; World Health Organization

OBJECTIVE: To evaluate the safety and efficacy of allogeneic natural killer (NK) cells in the treatment of primary hepatocellular carcinoma (HCC), and to elucidate the mechanism of NK cells therapy. METHODS: Twenty-one patients with primary HCC treated with allogeneic NK cells at the Fifth Medical Center of the PLA General Hospital were followed up for 1 year. Peripheral blood mononuclear cells (PBMCs) were isolated from patient-related donors and cultured in vitro for 15 days and infused to the patients in two consecutive days. Clinical data and laboratory data were collected and analyzed, including survival, clinical features, imaging changes, hematology, immunology, and biochemical indicators to evaluate the safety and efficacy of allogeneic NK cell therapy. The changes of peripheral blood lymphocyte subsets after treatment were also analyzed to explore the possible anti-tumor mechanisms. RESULTS: (1) Of the 21 patients with primary HCC, 11 patients were treated once, 5 patients were treated twice, and 5 patients were treated 3 times. After allogeneic NK cells infusion, 10 patients had fever, 1 patient had slight hepatalgia and 1 patient had slight headache, no other adverse events occurred including acute and chronic graft-versus-host disease (GVHD). They resolved spontaneously within 8 hours without other treatment. (2) The total disease control rate was 76.2% during one-year follow-up. Among them, the patients with Barcelona clinic liver cancer (BCLC) stage A had a disease control rate of 100%, stable disease (SD) in 10 cases; BCLC stage B patients had a disease control rate of 60%, partial response (PR) in 1 case, and SD 2 in cases; BCLC stage C patients had a disease control rate of 50%, complete response (CR) in 1 case, and 2 cases of PR. (3) The frequencies of NK cells and CD8+ T cells in peripheral blood were significantly lower than that before at 24 hours after treatment, and the frequencies of CD4+ T cells and CD4/CD8 were significantly higher than the baseline. CONCLUSION Allogeneic NK cells have good safety and efficacy in the treatment of primary HCC. The anti-tumor effect of the allogeneic NK cells may play an important role in the activation of the patient's natural immune system and delay disease progression, suggesting that allogeneic NK cells combined with sorafenib may be a very effective treatment for advanced HCC, and further large-sample multicenter randomized controlled clinical trials are needed to validate this result.
Carcinoma, Hepatocellular ; Graft vs Host Disease ; Humans ; Killer Cells, Natural ; Leukocytes, Mononuclear ; Liver Neoplasms