OBJECTIVETo investigate the apoptosis and proliferation effect of matrine on human medulloblastoma cell line D341 in vitro and the effect of the expression of the related caspase 3 and caspase 9 proteins.

METHODSThe D341 cells were cultivated successfully in vitro. Then the cells were divided into 5 groups according to the concentration of matrine (0.5 mg/mI group, 1.0 mg/ml group, 1.5 mg/ml group, 2.0 mg/ml group and the control group was 0 mg/ml). All the experiments were repeated three times. The cell morphologic and structure change was observed with the optical microscope and the transmission electron microscope. The proliferation of D341 cell was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining. The expression of Caspase3 and Caspase9 was detected by Western blot.

RESULTSWith the effect of matrine, the proliferation inhibition rate gradually increased with drug concentrations increasing, and there was a significant difference (P < 0.01). The inhibitory effect of matrine on cell proliferation was different with the different treatment time, there was a significant difference between the 24 h to 72 h groups (P < 0.01). The apoptotic rate increased with matrine concentrations increasing. There were significant differences between the group of 0.5 mg/mI or 1.0 mg/mI to the group of 1.5 mg/mI or 2.0 mg/mI (P < 0.05). The apoptotic rate increased with the prolonged treatment time. There were significant differences between the group of 24 h or 48 h to the group of 72 h ( P < 0.05). With the increase of matrine concentration, the expression of Caspase 3 and Caspase 9 increased (P < 0.01).

CONCLUSIONMatrine induces the apoptosis, and inhibits the proliferation of human medulloblastoma D341 cells in vitro by up-regulation of the expression level of Caspase3, Caspase9.

Alkaloids ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; metabolism ; pathology ; Humans ; Medulloblastoma ; metabolism ; pathology ; Quinolizines ; pharmacology ; Up-Regulation

160 s and core temperature was 36-37℃.Blood ga- ses,platelets,plasma free hemoglobin and coagulation factors were measured during ECMO support. The blood supply was monitored in the limbs cannulated with the femoral artery cannula.Results Two patients were successfully treated with ECMO without major complications.The circulatory and respiratory function of the patients was stable.The chest X-ray showed a no clouding of lungs and he- patic function was greatly improved in case 2 who underwent a successful heart transplantation follo- wing mechanical cardiocirculatory support as a bridge,Two patients received ECMO support for 5 and 3 days respectively.Both patients recovered well.No any severe acute rejection occurred and heart func- tion was NYHAⅠ.Conclusion When candidates suffered acute cardiogenic shock,ECMO can provide safe and effective mechanical circulatory support as a bridge to heart transplantation.

Objective To assess the value of multi-slice spiral CT angiography (MSCTA) in diagnosis of posterior nutcracker syndrome (NCS). Methods MSCTA of 15 patients clinically diagnosed as posterior NCS (patient group) and 50 subjects with normal renal vessels (control group) were retrospectively analyzed. The anatomy, course and relationship to the adjacent structure of left renal vein (LRV) and its branches were observed. The anteroposterior diameter (d1) and cross areas (s1) of the angle of control group and posterior abdominal artery (AA), the anteroposterior diameter (d2) and cross areas (s2) at the largest lumen near the renal hilar were measured and the ratio of s2/s1 and d2/ d1 were calculated.Results All LRVs posterior AA of patient group were compressed and narrowing with dilating apparently near the renal hilar. The difference of d2 was not significant (P＞0.05), but of other values were significant (P＜0.05).Conclusion MSCTA can show anatomy and three-dimentional trend clearly of LRV for posterior NCS, and evaluate the extent of narrow and dilating lumen accurately with cross areas measuring.

Objective To investigate the effect of different Chinese herbs on cell proliferation and cartilage oligomeric matrix protein(COMP)expression in chondrocyte culture.Methods Chondrocytes isolated from rabbit knee cartilage were cultured for 3 generations with the density of 2?10~4/cm~2 and were verified by collagenⅡimmunohistochemical staining.Rabbit sera containing herbs were obtained after animals orally ad- ministrated herbs at the dosage equivalent to human.At 5% and 10% serum density,cells were cultured in the medium that contained liver-softening herbal compound sera.Subgroups setting at 1,3 and 5 hours after herb intervention were observed.Rabbit and bovine sera were control groups.Seven days after intervention,chon- drocytes proliferation was observed using the MTT assay kit.For the study of COMP expression,chondrocytes were isolated from human knee cartilage supematant.Superuatant COMP level was tested by enzyme-linked immunoabsorbent assays(ELISA)after directly adding compound and extract from liver-softening herbs to the culture at the final concentration of 10 mg/ml for 3 days.Results Liver-softening herbal compound group had significant effect on cell proliferation compared to control,of which,3-hour subgroup was more significant than 1-and 5-hour subgroups(P

In order to investigate the mechanism, the correlation between the odor change in Crataegi Fructus stir-fried process and 5-HMF were studied. Required samples were retrieved from Crataegi Fructus stir-fried process. Statistical quality control (SQC) was used to analyze the response values acquired by the electronic nose. At the same time, the content of 5-HMF was detected by high performance liquid chromatography (HPLC). Correlation analysis was used to analyze the relationship between the above two. Experimental results showed that SQC model established by response values of all samples could show the change law of odor in Crataegi Fructus stir-fried process and changes of 5-HMF content was dropped after the first increase. Correlation analysis showed that the odor change in Crataegi Fructus stir-fried process and 5-HMF were significantly correlated (P < 0.05). Sugar degradation reaction and the Maillard reaction may be one of the mechanisms of the odor change in Crataegi Fructus stir-fried process.
Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Furaldehyde ; analogs & derivatives ; analysis ; Hot Temperature ; Odorants ; analysis ; Plant Extracts ; analysis ; Technology, Pharmaceutical ; methods

Anatomical features of the lamina and petiole of 11 species of Salvia had been compared. The microstructures of 11 species of Salvia were observed by paraffin method, and indexes such as the epidermis, xylems and phloems were measured. The data set was analyzed by principal components analysis method and cluster analysis method. The results indicated that the laminae of 11 species of Salvia were bifacial leaf, and the surface of laminae and petioles were covered with villous. The upper epidermis and lower epidermis of the laminae of 11 species of Salvia were constituted by a dense cuticular layer without stratum corneum. Differences existed in the structure of cross sectio mn of laminae and petioles of 11 species of Salvia. Among them, upper epidermal thickness of nine different micromorphological characters could serve as one of the criteria to identify species of taxonomic Salvia. The identification of genetic relationship of these characters in Salvia were discussed in the paper to provide an evidence of anatomy in Salvia.
Cluster Analysis ; Plant Leaves ; anatomy & histology ; Salvia ; anatomy & histology ; classification

Objective To investigate the mechanism and epidemiological characteristics of carbap-enem-resistant Proteus mirabilis ( PM) strains deficient in swarming motility. Methods PM strains were isolated from Hangzhou General Hospital of CAPF ( Chinese People′s Armed Police Forces) during January 2013 to December 2014. Bacterial motility and flagella of the PM strains were observed through semi-solid agar culture and flagella staining. Pulsed-field gel electrophoresis ( PFGE) was performed for homology anal-ysis. Antimicrobial susceptibility test and phenotypic confirmatory test were also carried out. PCR analysis and DNA sequencing were performed to confirm the genotype of resistant genes. Plasmid electroporation and S1-PFGE in combination with Southern blot hybridization were used to determine the location of the carbap-enem-resistant genes. Genetic structure of the blaKPC-2 gene was obtained by PCR mapping. Results A total of 42 PM isolates deficient in swarming motility were screened out and the resistance rates to imipenem and meropenem were 57. 1% and 52. 4%, respectively. PCR analysis and DNA sequencing confirmed that 24 carbapenem-resistant PM isolates deficient in swarming motility carried blaKPC-2 gene and belonged to three clones as indicated by the results of PFGE. Southern blot hybridization indicated that the blaKPC-2 gene was located on plasmids varying in size (26 kb, 55 kb and 139 kb). In addition, some of the strains harbored several resistant genes, such as blaTEM-1 , blaCTX-M-65 and rmtB. The genetic structures of strains carrying blaKPC-2 gene were ISKpn8, blaKPC-2 and ISKpn6-like from upstream to downstream. Conclusion Compared with the PM strains with swarming motility, the carbapenem-resistance rate was significantly higher in these PM strains deficient in swarming motility. Carbapenemases KPC-2 played an important role in the carbapen-em-resistant PM strains deficient in swarming motility. There was a cloning spread trend for carbapenem-re-sistant PM strains in our hospital. Clinicians should pay more attention to the risk of spreading.

Objective To explore the effect of transplantation of mesenchymal stem cells (MSCs) transfected with the human receptor activity modifying protein 1 (hRAMP1) gene on proliferation and apoptosis of vascular smooth muscle cells (VSMCs) after carotid balloon angioplasty was performed in rabbits with carotid atherosclerosis.Methods Density gradient centrifugation and adherent culture were carried out to obtain MSCs,which were then transinfected with an adenovirus vector carrying the hRAMP1 gene or an empty adenovirus vector.A rabbit model of atherosclerotic stenosis and balloon angioplasty was successfully established.Results were randomly divided into three groups:the hRAMP1-MSCs group,theadipose-derived MSCs (Ad-MSCs) group and the control group.MSCs were transinfected with Ad-EGFP-hRAMP1,Ad-EGFP or PBS by transplantation into the injured carotid arteries.Homing and differentiation were assessed with MSCs harvested at 7 d.With MSCs collected at 28 d,Western blotting was used to measure the expression of the hRAMP1 target gene in the carotid artery; the neointima and media area in the injured carotid arteries were estimated; carotid artery morphology was examined with H&E staining; and the proliferation and apoptosis of VSMCs were determined by immunohistochemistry and TUNEL.Results The expression of CD31 and EGFP was found in proliferating neointima lesions at 7d in the hRAMP1-MSCs group and the Ad-MSCs group.At 28d of MSC transplantation,the level of RAMP1 significantly increased in the hRAMP1-MSCs group,compared with the Ad-MSCs and control groups [(63.0±4.9) vs.(28.3±2.5) and (27.2±7.2),all P＜0.05],but there was no differencein the RAMP1 level between the Ad MSCs group and the control group (P＞0.05).Positive expression of the α-smooth muscle antibody (α-SMA) was found in all three groups at 28 d of MSC transplantation.The thickness of the hyperplastic neointima significantly decreased in the hRAMP1-MSCs group,compared with the other two groups (P＜0.05),and was lower in the Ad-MSCs group than in the control group (P＜0.05).The expression of proliferating cell nuclear antigen (PCNA) was lower in the hRAMP1-MSCs group than in the Ad-MSCs and control groups at 28d of MSC transplantation (P ＜0.05),while the PCNA level was lower in the Ad-MSCs group than in the control group (P＜ 0.05).The VSMC apoptosis rate significantly increased in the hRAMP1-MSCs group,compared with the Ad MSCs and control groups (P＜0.05),and was the lowest in the control group (P＜0.05).Conclusions Gene-modified stem cell therapy can effectively inhibit vascular intimal hyperplasia,thereby reducing restenosis after angioplasty.

Objective ① To Screen for the miRNAs differently expressed in the peripheral blood mono-nuclear cells (PBMCs) of rheumatoid arthritis (RA) by microarray experiments.② To further evaluate the expression of miR-155 in PBMCs of RA.③ To determine the relevance between the expression of miR-155 and clinical as well as laboratory features.④ To test whether inflammatory mediators can induce miR-155 expression in PBMCs of RA.Methods ① Total RNA was isolated from peripheral blood mononuclear cells obtained from 5 patients of RA and 5 normal controls.Expression profiling of miRNAs was performed in a microarray analysis.② MiR-155 was identified for further study by stem-loop real-time RT-PCR based on SYBR-Green.PBMC from 26 patients of RA and 23 normal controls were collected.③ Association between miR-155 and the clinical and laboratory features of RA was evaluated.④Induction of miR-155 following stimulation with TNF-α, IFN-γ and LPS of cultures of RA PBMCs was examined by real-time RT-PCR.Statistical analysis was done with student's t test, paired t test, and ANOVA, Spearman correlation.Results ① Expression profiling of miRNAs revealed significant differential expression of 14 miRNAs, of which signal intensity changed over two times.MiR-155 was up-regulated in PBMCs of RA than in normal controls (t=9.218, P=0.001).② The expression level of miR-155 had a positive correlation with serum CRP level (r=0.57, P=0.002).③ Expression of miR-155 was markedly up-regulated in PBMCs of RA after stimulated with TNF-α,IFN-γ and LPS, especially with TNF-α.Conclusions The expression of miR-155 is induced by stimulating with TNF-α, IFN-γ and LPS.MiR-155 may be a regulator in RA pathogenesis.Further studies are required to elucidate the function of miR-155.

Objective To screen and analyze nucleotide variants in 5'-untranslated region(5'-UTR)in voltage-gated sodium channel α1-subunit gene(SCN1A)in patients with Dravet syndrome and to evaluate the association of the variants with disease.Methods Peripheral blood of 24 patients with Dravet syndrome and 100 unrelated normal persons were collected and genomic DNA was extracted.PCR-sequencing of SCN1 A 5'-UTR in these DNA was performed.To evaluate the possibility of mutation inducing disease,bioinformatics analysis was applied to analyze the conservation of the sequences around the mutation site and predict the potential transcription elements.Results The nucleotide variant of 166.642.520G→A in exon 2 was identified in two patients,but not in normal controls.The mutation was a de novo mutation in a patient with early-onset.In the second proband,the mutation was also carried by his clinically asymptomatic mother.The nucleotide site 166.642.520 was moderately conserved in mammals(62.5%).The average nucleotide identity rate between human and other mammals species in the region adjacent to 166.642.520 was 88.5%.Two potential transcription regulatory elements were predicted on the sequence with the mutation of 166.642.520G>A,and only one on the sequence with wild-type.Conclusions The mutation 166.642.520G>A may be associated with Dravet syndrome and further studied should be performed to verify it and demonstrate its pathogenic mechanisms.