Ten glycosidic compounds were isolated from an ethanol extract of Machilus wangchiana by a combination of various chromatographic techniques including column chromatography over silica gel and Sephadex LH-20 and reversed-phase flash chromatography and HPLC. Their structures were identified by spectroscopic data analysis (IR, MS, and NMR) as icariside B1 (1), boscialin-3-O-β-D-glucopyranoside (2), pisumionoside (3), isolariciresinol-9'-O-β-D-xylopyranoside (4), 5'-methoxyisolariciresinol-9'-O-β-D-xylopyranoside (5), lyoniresinol-9'-O-β-D-xylopyranoside (6), (E) -4-hydroxyphenylprop-7-ene 4-O-β-D-glucopyranoside (7), (E) - 4-hydroxy-3-methoxyphenylprop-7-ene 4-O-α-L-rhamnopyranosyl-(1 --> 6) -β-D-glucopyranoside (8), 4-hydroxy-3-methoxyphenylprop-8-ene 4-O-β-D-xylopyraosyl-(1 --> 6) -β-D-glucopyranoside (9), and 4-hydroxy-3,5-dimethoxyphenylprop-8-ene 4-O-α-L-rhamnpyranosyl-(1 --> 6)-β-D- glucopyranoside (10), respectively.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Lauraceae ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo prepare monoclonal antibodies against oh(8)dG and to evaluate the relationship between Hp infection and oxidative DNA damage by detecting oh8dG in gastric mucosa.

METHODSBALB/C mice were immunized with BSA-oh(8)dG conjugate, monoclonal antibodies were prepared by hybridoma technique, the biological characteristics of antibodies were analysed by competitive ELISA, Western blot and immunohistochemistry.

RESULTSTwo strains of hybridoma cell were obtained. ELISA and Western blot indicated that the antibodies were fairly specific for oh(8)dG. In immunohistochemistry,the positive rate of oh(8)dG expression in Hp positive tissues and Hp negative tissues was 55% and 5%, respectively(P<0.01).

CONCLUSIONThe prepared antibodies can specially recognize oh(8)dG and immunohistochemistry with the monoclonal antibodies showed Hp infection can increase oh(8)dG level in gastric mucosa.

Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Deoxyguanosine ; analogs & derivatives ; analysis ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Gastric Mucosa ; chemistry ; Helicobacter Infections ; diagnosis ; Helicobacter pylori ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C

Objective To establish and evaluate criteria applied to review of complete blood counts (CBC)and differential results from automated hematology analyzers.Methods Temporary criteria were established by using alarm system of XE-2100 automated hematology analyzer and by consulting the 41 suggested rides of international consensus group.2 795 out-and in-patient samples were run as clinical samples.Stained blood films were prepared and manual differential with smear review were performed on all samples.Statistical analysis was done for each temporary rule and instrument flag which indicated abnormal cell quantity and morphology.Results Of all rules,instrument flags of ‘Immature Gran/Left Shift?’, ‘ Atypical Lympho?’and‘NRBC(nucleated red blood cell)?’showed most frequent false positive and false negative instrument flag.Evaluation on rnles about cell quantity change showed false positive and false negative rates were both low.Results of morphology evaluation showed that true positive rate was 17.44%, false positive rate was 15.82%,true negative rate was 63.49%,false negative rate was 3.25%.‘ Atypical Lymphocyte?’,‘Immature Gran?’and‘blast?’were the most frequent false positive flags.According to those results and clinicians opinions,our hematology review criteria for action following automated CBC and leukocytes differential was established.Conclusions The hematology review criteria have high true positive rate and low false negative rate.To clinical hematology laboratory using automated hematology analyzer,new criteria can reduce work load,bring lower false negative rate and higher work efficiency.

Female ; Hepatitis, Viral, Human ; therapy ; Humans ; Liver, Artificial ; Male ; Plasma Exchange ; methods ; Treatment Outcome

Objective To establish a rapid and accurate detecting method for sulfite in food residues. Methods Applying an fluorine ionsalkali - stable liquid to extract ultrasonically. After removing the suspended substances by high - speed centrifugation,we used online dialysis - ion chromatography to determine sulfite. Chromatographic conditionswere set as follows:Metrosep A Supp 5 - 150 / 4. 0 anion analysis column,using 3. 2 mmol / L sodium carbonate - 1. 0 mmol / L sodium bicarbonate 5% acetone as the eluent,with 0. 70 mL / min flow rate,20 μL injection volume and a conductivity detector. Results Sulfites( calculated as sulfur dioxide)manifested a good linear relationship when its concentration ranged from 0. 50 μg / mL to 50. 00 μg / mL,r = 0. 999 8. The minimum detection limit was 0. 54 mg / kg. The spiked concentrations were 10. 0 mg / kg,100. 0 mg / kg,500. 0 mg / kg,900. 0 mg / kg to samples. Relative standard deviation(n = 6,% )was 5. 04,1. 20,0. 56,0. 98,with the recovery rate between 85. 4% - 104. 4% . Conclusion This method is simple,rapid,with high sensitivity and accuracy of measurement,and which is suitable for the detection of sulfites content in food matrix.

OBJECTIVETo investigate the effects of damage-regulated autophagy modulator (DRAM) on radiosensitivity and the related mechanisms of implanted tumors of SGC7901 human gastric carcinoma cells in nude mice.

METHODSNude mice were randomly divided into model control group, radiotherapy group, and DRAM treatment group and radiotherapy combined with DRAM treatment group. When volume of transplantation tumor were 1.0 cm(3), radiotherapy, DRAM treatment was given. On days 3, 6 and 9 after treatment, the inhibition rate of tumor growth, pathological changes in tumor specimens, expression levels of P53, proliferating cell nuclear antigen(PCNA), C-myc, Fas-L, as well as apoptosis indexes in tumor samples were observed.

RESULTSInhibition rates of tumor in DRAM combined with radiotherapy were 9.3%, 14.1%, 16.7% on day 3, 6 and 9, respectively, all significantly higher than those in the radiotherapy group(5.0%, 8.8%, 6.5%, P<0.05). The expressions of PCNA and C-myc protein were down-regulated, while the expressions of P53 and Fas-L were upregulated.

CONCLUSIONDamage-regulated autophagy modulator gene may promote cell apoptosis and inhibit cell growth to enhance the radiosensitivity of transplanted gastric tumor in vivo in nude mice.

Animals ; Autophagy ; genetics ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Radiation Tolerance ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; radiotherapy ; Tumor Cells, Cultured

By using a combination of various chromatographic techniques including column chromatography over silica gel and Pharmadex LH-20 and reversed-phase HPLC, two minor new compounds, labda-12, 14-dien-6beta, 7alpha, 8beta, 17-tetraol (1), 2, 3-cis-6-acetyl-5-hydroxy-2-(hydroxymethylvinyl)-2, 3-dihydrobenzofuran-3-ol angelate (2), and a minor new natural product 6-methoxy-4-methyl-3, 4-dihydro-2H-naphthalen-1-one (3) have been isolated from an ethanolic extract of Heteroplexis micocephala. Their structures were elucidated with spectroscopic data analysis including 2D NMR experiments.
Aster Plant ; chemistry ; Benzofurans ; chemistry ; isolation & purification ; Diterpenes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Naphthalenes ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

BACKGROUNDIsocitrate lyase (ICL) was previously demonstrated to play a pivotal role in the intracellular metabolism of Mycobacterium tuberculosis (MTB). Presently several lines of evidence suggest that ICL from MTB (MTB-ICL) may play some roles in the interaction between MTB and host macrophage. However, there has been no research on the interaction between MTB-ICL and host macrophage.

METHODSMTB-icl and M. smegmatis (MS)-icl genes were amplified by polymerase chain reaction (PCR) and cloned into the E. coli-mycobacterium shuttle plasmid pUV15 to obtain recombinant shuttle plasmids pMTB-icl and pMS-icl. Following transformation into MS by electroporation, the expression of pMTB-icl and pMS-icl was verified by reverse transcriptase (RT)-PCR. The expression of recombinant plasmids derived from pUV15 when rMS was phagocytized by macrophage was also verified via fluorescence microscope. Ms 1 - 2c, rMS-pUV15, rMS-pMS-icl and rMS-pMTB-icl were used to infect RAW264.7 cells and the survival of intracellular MS was monitored by bacterial culture at 0, 24 and 48 hours after infection. The culture supernatants from macrophage infected by Ms 1 - 2c, rMS-pUV15, rMS-pMS-icl and rMS-pMTB-icl were collected and the interferon (IFN)-gamma and nitric oxide (NO) concentrations were measured by ELISA or by Griess assay, respectively. The apoptosis of macrophage was assayed by the in situ TUNEL technique.

RESULTSRT-PCR showed that both pMTB-icl and pMS-icl could be expressed in MS. Fluorescence microscopic observation showed that recombinant plasmids derived from pUV15 (pUV15-IG) could also be expressed in MS when MS were phagocytized by macrophage. Bacterial culture data demonstrated that rMS-pMTB-icl exhibited significantly increased intracellular survival in the murine macrophage cell line RAW264.7 compared with Ms 1 - 2c, rMS-pUV15 and rMS-pMS-icl. This increased intracellular survival was not accompanied by the upregulation of IFN-gamma and NO in host macrophage. But a lower apoptosis rate of macrophages infected with rMS-pMTB-icl was observed when compared with macrophages infected with other strains of MS.

CONCLUSIONSMTB-ICL could promote the intracellular survival of MS. Suppressing the apoptosis of host macrophage may be one of the important mechanisms involved in this increased intracellular survival.

Animals ; Apoptosis ; genetics ; physiology ; Cell Line ; In Situ Nick-End Labeling ; Interferon-gamma ; metabolism ; Isocitrate Lyase ; genetics ; metabolism ; Macrophages ; cytology ; metabolism ; microbiology ; Microbial Viability ; Microscopy, Fluorescence ; Mycobacterium smegmatis ; enzymology ; genetics ; growth & development ; Mycobacterium tuberculosis ; enzymology ; genetics ; Nitric Oxide ; metabolism ; Plasmids ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transformation, Genetic

This study was purposed to investigate the effect of highway transportation on the quality of blood components so as to provide experimental basis to meet the needs of military operations. The transport condition was simulated by random vibration test. The red blood cells, leukocyte-reduced red blood cells, washed red blood cells were randomly vibrated (C Road) for 4 hours. Then, these blood components were stored in refrigerator for 15 days (4 degrees C). Six milliliters of blood were collected before vibration, after vibration, and at day 15 days of storage after vibration, respectively. The suspension was isolated. The free hemoglobin (FHb), routine hematological parameters, and biochemical indexes were determined. The results showed that FHb, lactate dehydrogenase (LDH), K(+) of red blood cells and leukocyte-reduced red blood cells did not significantly change after vibration and storage. However, FHb, LDH and K(+) of washed red blood cells increased significantly after simulated transportation (p < 0.05). The levels of these parameters at day 15 of storage after vibration were also significantly higher than those after vibration (p < 0.01). The changes of other hematological parameters were not significant in three blood components after vibration (C Road) and storage for 15 day. In conclusion, red blood cells and leukocyte-reduced red blood cells were qualified for clinic transfusion even after transportation within 4 hours for 15 day storage later, if they were kept in proper blood container and protected from damping. However, the washed red blood cells could not be used for clinic after similar transport in the military operations.
Blood Preservation ; Cryopreservation ; Erythrocytes ; chemistry ; Humans ; L-Lactate Dehydrogenase ; blood ; Transportation ; Vibration

AIDS-Related Opportunistic Infections ; diagnostic imaging ; drug therapy ; etiology ; Adult ; Bone Marrow Examination ; CD4 Lymphocyte Count ; Female ; Humans ; Male ; Middle Aged ; Mycoses ; diagnostic imaging ; drug therapy ; etiology ; Penicillium ; isolation & purification ; Prognosis ; Radiography