Genomic DNA of two fungi Thamnidium elegans and Umbelopsis isabellina were extracted with an amended Cetyltrimethyl Ammonium Bromide (CTAB) method. This modified method uses repeated freezing in liquid nitrogen and thawing with combination of shocking with glass beads to replace of the tra- ditional method. Quality and concentration of DNA extracted by the modified methodwere tested. Compared with the traditional method, higher yield and purity of genomic DNA were obtained with less amount ofmy- celium. The result indicted that this is a simple and highly efficient method, which is suitable to treat many samples at one time and for basic molecular experiments, such as restriction endonuclease reaction and PCR.

OBJECTIVETo investigate the expression of PCNA and p27 in human benign prostate hypertrophy (BPH) and prostate carcinoma (PCa) and their effect on the genesis and progression of the tumor.

METHODSThe paraffin-embedded sections of 30 cases with BPH and 37 cases with PCa were collected. The expression of p27 and PCNA protein were examined by S-P immunohistochemical method. Comparative analysis for BPH and pathological grade and clinical stage of PCa was performed.

RESULTSThe expression of PCNA in BPH (3.3%) was significantly lower than that in Pca (83.8%, P < 0.01). The expression of p27 in BPH (70.0%) was significantly higher than that in Pca (27.0%, P < 0.05). The expression of p27 was not correlated with histological grade and clinical stage in Pca (P > 0.05). An inverse correlation was found between p27 and PCNA expression in BPH (P < 0.01), while no correlation was found in Pca (P > 0.05).

CONCLUSIONThe loss or decreased expression of p27 protein may be related to the genesis of benign prostate hypertrophy, but not to the development of prostate carcinoma; the overexpression of PCNA may play an important role in the malignant behavior and progression of prostate carcinoma.

Adenocarcinoma ; metabolism ; pathology ; Aged ; Aged, 80 and over ; Cell Cycle Proteins ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Proliferating Cell Nuclear Antigen ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Tumor Suppressor Proteins ; metabolism

AIM To study the chemical constituents from Pterocephalus hookeri (C.B.Clarke) Hoeck and their neuroprotection activities.METHODS The 95％ ethanol extract from P.hookeri was isolated and purified by Sephadex LH-20,DM-130 macroporous resin,silica and HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The neuroprotection activities were studied by establishment of Parkinson's disease model of genetically modified zebrafish.RESULTS Nine compounds were isolated and identified as triplostoside A (1),cantleyoside (2),sylvestroside Ⅰ (3),sylvestroside Ⅲ dimethyl acetal (4),sylvestroside Ⅲ (5),laciniatoside Ⅰ-7-dibutyl acetal (6),laciniatoside Ⅰ (7),laciniatoside Ⅱ (8),sylvestroside Ⅳ (9).The n-BuOH fraction of P.hookeri had good performance on neuroprotection in hydrogen peroxide model.CONCLUSION Compound 4 is isolated from this plant for the first time.P.hookeri has neuroprotection activities.

OBJECTIVETo analyze the differences in the immediate effect of acupuncture on cervical spondylosis of vertebral artery type among three factors: needling technique, acupoint selection and time of needle retaining so as to option the best therapeutic program.

METHODSThirty-two cases were observed and randomly divided into 8 groups, 4 cases in each one. The orthogonal design of three factors and two levels was adopted. There were needling technique (electroacupuncture, reinforcing and reducing method), acupoint selection [C4-C7 Jiaji (EX-B 2) and three needles of neck: Tianzhu (BL 10), Jingbailao (EX-LHN 15), Dazhu (BL 11)] and time of needle retaining (5 min, 15 min). L8 (2(7)) orthogonal design table was arranged in the trial. The changes in mean velocity (Vm) of vertebral artery (VA) and basilar artery (BA) were observed before and after acupuncture.

RESULTSThe immediate effect of VA-BA blood flow was the most significant after electroacupuncture at C4-C7 Jiaji (EX-B 2), with continuous wave for 5 min. This method and acupoint selection greatly influenced the therapeutic results (both P<0.05).

CONCLUSIONThe optimized therapeutic program of the immediate effect of acupuncture for cervical spondylosis of vertebral artery type is electroacupuncture + C4-C7 Jiaji (EX-B 2) + 5 min. The importance of different factors for the immediate effect in acupuncture treatment of cervical spondylosis of vertebral artery type is: acupoint selection > needling technique > time of needle retaining.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Regional Blood Flow ; Spondylosis ; physiopathology ; therapy ; Vertebral Artery ; physiopathology ; Young Adult

Aneurysm ; pathology ; Aneurysm, Dissecting ; diagnosis ; Cardiovascular Abnormalities ; pathology ; Deglutition Disorders ; pathology ; Humans ; Male ; Middle Aged ; Subclavian Artery ; abnormalities ; pathology

Animals ; Cell Differentiation ; Female ; Hematopoietic Stem Cells ; cytology ; Hepatocytes ; cytology ; Male ; Mice ; Mice, Inbred BALB C

OBJECTIVETo investigate the changes of mandibular vitodynamics in powerful bite working condition and impact working condition after mandibular angle osteotomy through 3-dimensional finit element analysis.

METHODSA patient of prominent mandibualr angle without malocclusion was selected and underwent 3-dimensional CT before and after operation. The DICOM data of 3-dimensional CT were read by Mimics software and 3-dimensional images were reconstructed. Then the 3-dimensional images were changed into IGES format and imported into ANSYS10.0. The boundary constrained condition and pre-processing condition was setting in ANSYS10.0 software. The 3-dimensional finite element models were generated in ANSYS10.0. Solution process was running and most powerful bite working condition and impact working condition were simulated and calculated.

RESULTSIn static stress analysis, the stress peak and stress maximal value point of mandible were basically at equal pace in preoperative and postoperative models. In transience dynamic stress analysis, although stress discrepancies were found in part time point and some region of mandible between preoperative model and postoperative model, the direction and topography of main stress were basically at equal pace. Main stress distributed beside external oblique line and concentrates at the neck of condyle. There was no significant difference of stress conduction and distribution between preoperative and postoperative models.

CONCLUSIONSCompared with preoperative model, postoperative model has different transduction phases of stress, but has almost the same tolerance extents of main stress.

Female ; Finite Element Analysis ; Humans ; Imaging, Three-Dimensional ; methods ; Mandible ; physiopathology ; surgery ; Osteotomy ; Stress, Mechanical

Pseudomonas aeruginosa is an important opportunistic human pathogen. It encodes many virulence factors and one of them is type III secretion system (TTSS). Effectors proteins can be delivered into host cells directly by this system, causing necrosis or apoptosis. popN gene is the first gene in the popN operon of TTSS gene cluster. To investigate its function, popN gene deletion mutant was generated in this study, and we found this mutant can secrete effectors proteins constitutively under non-inducting condition in DMEM medium containing serum. The results indicated that PopN is a negative regulator of the TTSS expression. However, no secreted effector proteins were detectable when the popN- mutant was grown in LB medium under non-inducting condition. To investigate the possible reasons, effects of growth status and protease (s) inhibitors on the TTSS were investigated. We present evidences that indicate protease mediated degradation of secreted effector proteins played a key role in the phenotypic inconsistency of popN- mutant.
ADP Ribose Transferases ; metabolism ; secretion ; Bacterial Proteins ; genetics ; metabolism ; secretion ; Bacterial Toxins ; metabolism ; Gene Expression Regulation, Bacterial ; Mutation ; Peptide Hydrolases ; genetics ; metabolism ; Pore Forming Cytotoxic Proteins ; genetics ; secretion ; Protease Inhibitors ; pharmacology ; Pseudomonas aeruginosa ; genetics ; metabolism ; pathogenicity

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

OBJECTIVETo investigate the correlation of the expression of VEGF-C and VEGFR-3 to the pathological grade of human prostate cancer.

METHODSImmunohistochemistry was used to detect the expression of VEGF-C and VEGFR-3 in 25 cases of prostate cancer tissues.

RESULTSThe total positivity rates of VEGF-C and VEGFR-3 were 80% and 76% in these cancer tissues, respectively. The positivity rates of VEGF-C was 94.7% in the 19 cases with Gleason scores no less than 6 (group I), significantly higher than the rate (33%) in the 6 cases with Gleason scores between 4 and 6 (group II) (P<0.01). The positivity rates for VEGFR-3 also showed a significant difference between groups I and II (89.5% vs 33.3%, P<0.05). The expression level of VEGF-C was correlated to the Gleason score of prostate cancer (R=0.436, P<0.05), and the correlation between VEGFR-3 and the Gleason score was even more obvious (R=0.608, P<0.01). Their expressions, however, did not show any correlations to the patients age, PSA or the volume of the prostate.

CONCLUSIONVEGF-C and VEGFR-3 may serve as new markers for evaluating the malignancy of prostate cancer with Gleason score not less than 4.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Neoplasm Grading ; Prostatic Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor Receptor-3 ; metabolism