[Objective] To investigate the expression of MICA mRNA and protein in osteosarcoma and give a more comprehensive understanding to its immune evasion.[Methods] RT-PCR was used to analyze MICA mRNA in 11 osteosarcoma tissues and 5 osteosarcoma cell lines.MICA expression was examined in 66 paraffin-embedded osteosarcoma tissues and 6 paraffin-embedded normal bone tissues by immunohistochemistry,and MICA protein in 9 fresh osteosarcoma tissues was detected by Western blot too.MICA surface expression was estimated by flow cytometry.[Results] Nine of eleven (81.8%) osteosarcoma specimens and all of the five cell lines consistently expressed MICA mRNA.Up-regulation of MICA expression was found in osteosarcoma (34/66,51.6%),compared with normal bone tissues (0/6,0%).The cell lines Saos-2,MG63,and HOS showed positive surface MICA expression,while the U2OS and OS732 showed very limited expression.[Conclusion] MICA mRNA and protein predominantly expressed on osteosarcoma tissues and cell lines.

Adolescent ; Diagnosis, Differential ; Humans ; Male ; Sturge-Weber Syndrome ; pathology

Humans ; Lung Neoplasms ; metabolism ; pathology ; surgery ; Lymph Node Excision ; Male ; Middle Aged ; Pneumonectomy ; methods ; Pulmonary Blastoma ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

This paper is aimed to microscopic identification of traditional Chinese medicines (TCMs) using an in situ imaging method. In this study, two kinds of Zingiberaceae seeds, Amomi Rotundus Fructus and Alpiniae Katsumadai Semen, were investigated by synchrotron radiation in-line X-ray phase-contrast computed tomography (IXPCT) imaging method. The results showed that the microstructures of these Zingiberaceae seeds could be clearly obtained from the virtual slices information in different observing angles. It proves that IXPCT is an effective imaging method, which can provide the imaging information for the microscopic identification of the intact TCMs in situ and non-destructively.
Amomum ; cytology ; Imaging, Three-Dimensional ; methods ; Medicine, Chinese Traditional ; Seeds ; cytology ; Tomography, X-Ray Computed

In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.
China ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; classification ; standards ; Geography ; Marsdenia ; chemistry ; classification ; Medicine, Chinese Traditional ; Principal Component Analysis ; Quality Control ; Reproducibility of Results ; Spectroscopy, Fourier Transform Infrared ; methods

This study was aimed to identify the main related substance in baicalein in order to provide basic study data for the safety, efficacy and quality control of the medicine. The analysis was carried out on activated carbon and silica gel column. The compound structure was identified by spectral analysis, which included UV, IR, NMR and MS. The results showed that the related substance was identified as oroxylin A. It was concluded that the determina-tion provided scientific data for studying the impurities of baicalein and improving its quality.

To offer the reference and method for salt damage in the cultivation of Marsdenia tenacissima, the seeds of M. tenacissima collected from Maguan city ( Yunnan province) were taken as the test materials to study the effects of different priming materials on improving germination and growth under high-level salt stress condition. Four different treatments, which were GA3, KNO3-KH2PO4, PEG-6000, NaCl, combined with ANOVA were applied to test the performance of germination energy, germination percentage, germination index, MDA, SOD, and CAT. The results showed that the seed germination was obviously inhibited under salt stress and the soaked seeds with different priming materials could alleviate the damage of salt stress. Under these treatments, the activities of SOD, CAT the content of soluble protein significantly increased. While the content of MDA significantly decreased. The maximum index was obtained when treated with 1.20% KNO3-KH2PO4, the germination percentage increased from 52.67% to 87.33% and the activity of SOD increased from 138.01 to 219.44 respectively. Comparing with the treatment of 1.20% KNO3-KH2PO4, the germination percentage of treating with 300 mg x L(-1) GA3 increased from 52.67% to 80.67%, while the activity of SOD increased from 138.01 to 444.61.
Germination ; drug effects ; physiology ; Marsdenia ; drug effects ; growth & development ; Nitrates ; pharmacology ; Polyethylene Glycols ; pharmacology ; Potassium Compounds ; pharmacology ; Seeds ; drug effects ; growth & development ; Sodium Chloride ; pharmacology ; Stress, Physiological ; Xanthones ; pharmacology

OBJECTIVETo express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.

METHODSHA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.

RESULTSWe successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.

CONCLUSIONWe expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.

Animals ; Blotting, Western ; Cell Line ; Drosophila ; Female ; Gene Expression ; Hemagglutinin Glycoproteins, Influenza Virus ; analysis ; chemistry ; genetics ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; metabolism ; Influenza, Human ; virology ; Mice ; Mice, Inbred BALB C ; Solubility

Tyrosol, crenulatin and salidroside are the main active constituents of Rhodiola crenulata, with extensive pharmacological activities. In the study, grams of high purity tyrosol, crenulatin and salidroside were simultaneously separated from R. crenulata by the first time. Firstly, R. crenulata was extracted by 70% alcohol. Then, with the yields of three compounds as the index, the macroporous resin was optimized. At last, grams of high purity tyrosol, crenulatin and salidroside were isolated by D-101 macroporousresin, purified by column chromatography. Detected by HPLC, the purity of three compounds were higher than 98%. This method has the advantages of simple process and operation, less dosage of organic solvent, highly yield and reproducibility, suitable for the simultaneously preparation of tyrosol, crenulatin and salidroside.
Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; Coumarins ; analysis ; isolation & purification ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Glucosides ; analysis ; isolation & purification ; Phenols ; analysis ; isolation & purification ; Phenylethyl Alcohol ; analogs & derivatives ; analysis ; isolation & purification ; Rhodiola ; chemistry

OBJECTIVETo study the expression of membrane MICA (mMICA), soluble MICA (sMICA) and NKG2D receptor in cases of osteosarcoma and to analyze its clinical significance.

METHODSExpression of mMICA in osteosarcoma tissue of 43 cases was detected with immunohistochemistry. Expression of NKG2D in peripheral blood lymphocytes of 16 cases was analyzed by flow cytometry. Serum level of soluble MICA (sMICA) was measured by ELISA.

RESULTSmMICA was widely expressed in osteosarcoma tissue (37/43). Expression of NKG2D in peripheral blood lymphocytes was significantly decreased. High levels of mMICA and NKG2D expression were associated with better differentiation and earlier tumor stage of osteosarcoma (P < 0.05). A significant increase in serum level of sMICA was demonstrated in patients with metastasis and advanced tumor.

CONCLUSIONSThe mMICA expression in tumor tissue, NKG2D expression in peripheral lymphocytes and serum sMICA level correlate with the differentiation and stage of osteosarcoma. These parameters may thus represent potential diagnostic and prognostic markers in patients with osteosarcoma. Manipulation of the MICA-NKG2D pathway may become a target of immunotherapy for osteosarcoma.

Bone Neoplasms ; blood ; metabolism ; pathology ; Cell Differentiation ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Histocompatibility Antigens Class I ; blood ; metabolism ; Humans ; Immunohistochemistry ; Lymphocytes ; immunology ; NK Cell Lectin-Like Receptor Subfamily K ; metabolism ; Neoplasm Staging ; Osteosarcoma ; blood ; metabolism ; pathology