Animals ; Eye Diseases ; parasitology ; surgery ; Humans ; Middle Aged ; Sparganosis ; diagnosis ; surgery ; Sparganum

Sixteen compounds were isolated from the ethanol extract of Illigera rhodantha by silica gel, ODS, and Sephadex LH-20 column chromatographies. These compounds were identified as (2R,3R)-2,3-dihydroxy-2-methylbutane-1,4-diyldibenzoate (1), p-hydroxyphenethyl trans-ferulate (2), 4-O-benzoyl-2-C-methyl-D-erythritol (3), N-trans feruloyl-3-methyldopamine (4), tribulusamide A (5), cryptomeridiol (6), teuclatriol (7), oleanolic acid (8), icario A₂ (9), vanillic acid (10), p-hydroxybenzoic acid (11), gallic acid (12), ethyl gallate (13), chrysophanol (14), D-1-O-methyl-inositol (15), and hexadecanoic acid (16) based on their spectral data and physico-chemical properties. Compound 1 is an undescribed compound, of which its absolute configurations were determined by Mosher and ROESY methods; all the compounds except 10, 11 and 14 were isolated from Illigera genus for the first time. Compared with the positive control indomethacin, compounds 4-6, 8 and 9 inhibited significantly against the NO production in LPS-induced RAW 264.7 cells.

Objective To study the application and significance of constructivism theory in pathological experiment teaching.Methods Six classes in total of 198 students majoring in clinical medicine were selected from grade 2009 of Guangdong medical college for research and were divided into experimental group (n=99) and control group (n=99) using random number table.Various teaching methods based on constructivism theory were used in experimental group while conventional teaching methods were applied in control group.The observing ability for specimen and slices,theoretical level and learning behavior of students were assessed with the same standard and anonymous homemade questionnaire survey was executed for students.SPSS 13.0 statistical software was used to analyze the results.The examination results of the two groups were compared using independent samples t-test.The questionnaire survey results of the two groups were compared using Wilcoxon rank-sum test.P＜0.05 represented the difference having statistical significance.Results The observing capability for specimen and slices,theoretical level and learning behavior of students in experimental group were significantly improved (P＜0.01)and the questionnaire survey results demonstrated that there were statistical difference in feedbacks between the two groups and the satisfaction rate was significantly improved in experiment group (P＜0.01).Conclusions Applying comprehensive teaching methods based on constructivism theory can stimulate students' interest and initiative in learning,improve students' learning behavior and learning strategies,and significantly improve the teaching quality.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD56 Antigen ; metabolism ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Fever of Unknown Origin ; etiology ; Humans ; Lymphoma, Extranodal NK-T-Cell ; complications ; metabolism ; pathology ; therapy ; Male ; Prednisone ; therapeutic use ; Splenectomy ; Splenic Neoplasms ; complications ; metabolism ; pathology ; therapy ; Vincristine ; therapeutic use ; Young Adult

OBJECTIVETo investigate the effects of ligustrazine on nitric oxide (NO), malonaldehyde (MDA) production, release of intracellular lactate dehydrogenase (LDH) and membrane fluidity of the injured human umbilical vein vascular endothelial cell line (ECV-304) with hypoxia and lack of glucose.

METHODThe experiments were performed in culture of ECV-304 injured with hypoxia and lack of glucose in vitro. The released LDH of ECV-304 was measured with automatic biochemistry analyse. NO content of ECV-304 was monitored with colorimetry. Lipid peroxidation of ECV-304 was monitored as MDA with a fluorometric assay. The membrane fluidity of ECV-304 was measured with the fluorescence polarization method.

RESULTAfter culture ECV-304 in hypoxia and lack of glucose for 24 h, the LDH release, MDA production and the membrane fluidity increased significantly and NO level was decreased. Preincubation of ECV-304 with ligustrazine for 24 h reduced LDH release, MDA production, membrane fluidity increasing and increased the level of NO in ECV-304 due to hypoxia and lack of glucose.

CONCLUSIONLigustrazine has protective effect on injury of ECV-304 induced by hypoxia and lack of glucose.

Cell Hypoxia ; Cell Line ; Cell Membrane ; drug effects ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Hypoglycemia ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Ligusticum ; chemistry ; Malondialdehyde ; metabolism ; Membrane Fluidity ; drug effects ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Protective Agents ; pharmacology ; Pyrazines ; isolation & purification ; pharmacology

Objective To investigate genotyps of Treponema pallidum (Tp) in several cities in Guangxi province. Methods A total of 300 patients with suspected early syphilis were enrolled from STD clinics in Guangxi between January 2012 and July 2016, and tissue fluid samples were collected from skin lesions. Silver staining was performed to detect Tp, and PCR to amplify the Tp polA gene for the diagnosis of early syphilis. Positive samples were subjected to PCR amplification of a 60-bp tandem repeat region within the arp gene, restriction fragment length polymorphism(RFLP)analysis of the tpr Ⅱgene after digestion with Mse Ⅰ enzyme and tp0548 genotyping. Results Finally, 215 patients were diagnosed with early syphilis, including 210(97.7%)patients positive for PCR and 105(48.8%)patients positive for silver staining, and the positive rate significantly differed between the two methods (χ2 = 103.01, P < 0.05). Among the PCR-positive samples, 190 could be genotyped by analysis of three target genes, and 17 genotypes were identified. The genotype 14d/f was predominant (45.3%, 86/190), followed by 15d/f (13.7%, 26/190), 16d/f(11.6%, 22/190), 17d/f(7.4%, 14/190), 13d/f(6.8%, 13/190), 10d/f(4.2%, 8/190), 18d/f(1.6%, 3/190), 16a/f(1.6%, 3/190), 5d/f(1.1%, 2/190), 7d/f(1.1%, 2/190), 12d/f(1.1%, 2/190), 16d/e(1.1%, 2/190), 14a/f(1.1%, 2/190), 9h/c(1.1%, 2/190), 15l/f(0.5%, 1/190), 25a/e(0.5%, 1/190), 15i/f(0.5%, 1/190). Conclusion Tp genotypes are diversified in patients with early syphilis in Guangxi, and the genotype 14 d/f is predominant.

Objective To study the characteristics of a new neuron-specific marker in the cerebral cortex and its application method.Methods According to immunofluorescence principles,the neurons in the cerebral cortex cultured in vitro were stained by fluorescein isothiocyanate (FITC)-labeled Tet1; and FITC-Tet1 with NSE or GFAP was employed to determine whether the neurons can be marked and whether they can be distinguished from astrocytes; the conditions of application of FITC-Tet1 were quantified.The HEK and C6 cell lines were performed the above experiments as controls.Results The final concentration was the key factor,which indicated that when the FITC-Tet1 was about 30 μg/mL,the neuron marking ability was the best,and the astrocytes,HEK and C6 were not stained.Neither high (100μg/mL) or low (10μg/mL) levels of FITC-Tet1 were suitableforstainingneurons.Conclusion Tet1 can specifically combine with the neuron in the cerebral cortex,which may make it to be a new staining method or a targeting molecule to neurons in the central nervous system.

Animals ; Cromolyn Sodium ; administration & dosage ; Disease Models, Animal ; Intestines ; blood supply ; Mast Cells ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; mortality ; pathology ; p-Methoxy-N-methylphenethylamine ; administration & dosage

The present paper is aimed to explore the biological osteoinductive activity of recombinant human bone morphogenetic protein 9 (rhBMP-9) by various biological technologies. In this study, we firstly obtained hBMP-9 cDNA by PCR and inserted it into vector pcDNA4/His Max to reconstruct hBMP-9 eukaryotic expression vector pcDNA4/His Max-BMP-9. Recombinant Chinese hamster ovary (rCHO) cell line expressing high-level rhBMP-9 was reconstructed by co-transfecting the expression vectors pcDNA4/His* Max-hBMP-9 and plasmid pSV2-dhfr into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification by the methotrexate. We finally obtained a monoclonal cell line expressing the highest level protein. We purified the medium after culturing the highest-producing monoclonal by Ni-NTA His-Bind Resin columns and concentrated to by a Centricon 50 at 4 degrees C and stored at 70 degrees C until it was used. Western blot and SDS-PAGE analyses showed a specific band of about 32kD in pro-region lane and a specific band of about 50kD in pro-region complex lane. Biological activities of rhBMP-9 were tested by colorimetric determination and histochemical staining of Alkaline Phosphatase (ALP) Activity, osteocalcin and oesteopontin for C3H10 T1/2 cells, which were stimulated culture by different concentration (20, 50, 100 microg/mL) of rhBMP-9. The results showed that the rhBMP-9 could induce osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro, and were proportional to the amount. This study can provide experimental data for further tests in vivo and clinical applications.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Growth Differentiation Factor 2 ; biosynthesis ; isolation & purification ; pharmacology ; Humans ; Osteogenesis ; drug effects ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology ; Transfection

Objective The aim of this study was to investigate the aquaporin-4(AQP4) expression in the ischemic penumbra tissues.Methods Thirty-six Wistar rats were divided into 7 groups randomly, including control group(n=6) and occluded groups(n=30). The occluded groups were studied after the right middle cerebral artery of the rats unilaterally occluded(MCAO) at an interval of 15 min, 30 min, 1 h, 3 h, 6 h and 24 h, respectively(n=5 for each group). The operation process of the control group was the same as the occluded group except occluded MCAO. Then all rats were imaged with T_1WI, T_2WI and diffusion weighted-imaging(DWI). The brain tissue, according to the method by LIU Meili reported, was regarded as the area of the graphic penumbra. The relative apparent diffusion coefficient of the graphic-penumbra (rADC_1) and the center infarction(rADC_2)(ratios between the values of the occluded side and the opposite side) were calculated. The animals were sacrificed and perfused with the mixture solution consisting of TTC at different time intervals. The graphic-penumbra of the biggest layer of the ischemic cerebral tissue which corresponded to the DWI was examined with immunohistochemistry and RT-PCR. Meanwhile, histologic examination was performed at same site of the lesion. Results There were no significant changes on MRI, the relative apparent diffusion coefficient and the expression of the AQP4. The abnormal high intensity was found on DWI at 15 min after MCAO. T_2WI detected the lesion at 1 h after MCAO. The value of the rADC_1 decreased within 24 h after MCAO in ischemic penumbra, especially, it descended quickly within 1 h after MCAO, from(70.4?6.9)% at 15 min to(53.5?10.9)% at 1 h. Whereas, in the infarct tissue, the changes of the rADC_2 had a rule of decrease from(71.5?6.6)% at 15 min to(45.7?10.5)% at 3 h at first time, and then follow an increasing up to(78.7?11.5)% at 24 h after MCAO. The expression of AQP4 increased gradually within 24 h after MCAO, from 0.42?0.05 at 15 min to 1.18?0.12 at 24 h, it showed negative relationship with the rADC_1 in the ischemic penumbra (r= -0.966,P