A bacterium that contains a chitin deacetylase has been isolated from ocean soil. It has been identified Baciluus .The best suitable condition of producing this enzyme is that pH is 4.0,35℃,and the incubate time is 80 hours in the present of Ca 2+ as well as having no chitosan. The best temperature of this enzyme when it acts is 40℃～50℃ and the best pH range is 4.5～5.0.

Heparinases,a kind of polysaccharide lyases,can degrade heparin and heparin sulfate to low molecular weight polysaccharides.It has been noted that many bacteria have heparinases although only few of them have been purified and characterized.Heparinases I,II and III from Flavobacterium heparinum have been extensively studied for many years and been commercialized recently.Heparinases have some important applications in the industry and clinic as well as in the determination of heparin structure,which is a very important anticoagulant drug used world-widely.The recent progresses in isolation of heparinase-producing bacteria,genome mapping of heparinase homologs in sequenced bacteria and archaea genomes,purification of heparinases and the study of their biochemistry and regulation were reviewed.The recombinant expression of these enzymes as well as important applications of heparinases and their potential applications in the future will also be highlighted.

AIM: To identify the non-steroid transcription factors upregulating the expression of L-plastin in hormone-independent prostate cancer, and partly elucidate the mechanism of hormone-refractory prostate cancer. METHODS: TF SEARCH software was used to analysis the possible binding sites of transcription factors in the 3’ end of L-plastin promoter that had been identified as important part of regulation response elements. Gel shift assay and supershift assay were used to confirm the transcription factors binding the speculated response elements. PCR site-mutagenesis technique was performed to delete the binding site of transcription factor and luciferase activity assay was carried out after deletion of the binding site. RESULTS: SP-1 respond element GGTGGGGCGGGGA located at -54- -41 of L-plastin promoter was identified with the TF SEARCH software. Gel shift assay and supershift assay confirmed that SP-1 was the transcription factor binding to GGTGGGGCGGGGA. Mutant deleted the SP-1 binding-site had low-luciferase activity than that of the naive. CONCLUSION: SP-1 plays an important role in the up-regulation of L-plastin expression in hormone-independent prostate cancer.

Vitamin C precusor-2-keto-L-gulonic acid can be prepared directly by mixed culture of Ghiconobacter oxy-dans SCB329 and Guconobacter subaxydans SCB110. To obtained its high yield, firstly, the proportion of the two micro- organisms, the ingredients of medium and the initial pH were optimized in shake flaskd, then L9 (34) orthogonal experiment confirmed that urea, C. S. L had high degree statistical meaning. Based on these data, an optimized fermentation media was obta ined: D-Sorbitol 9g, C. S.L1.5g, Urea1.5g, KH2PO40.1g, CaCO30.2g. By-product can be inhabited to the greatest extent and the yield increases by 20%.

OBJECTIVETo investigate the MRI diagnosis of bone contusion on the knee, and improve the diagnosis skill.

METHODSA retrospective analysis of 47 cases of knee bone bruise with the performance of MRI in our hospital from 2008.5 to 2009.12, including 30 males and 17 females,aged from 12 to 69 years, with a mean of 34.5 years old. Forty-seven patients suffered from post-traumatic localized pain, tenderness and(or) soft tissue swelling and dysfunction. The time interval between the accident and the MRI examination ranged from 6 h to 30 days, averaged 7 days. The patients with only bone contusion were treated with conservative and symptomatic treatment.

RESULTSA total of 47 cases showed 82 lesions, including 39 distal femur, 35 proximal tibias (including the tibial plateau),5 patellas, 3 fibula top. MRI showed irregular patchy or map-like T1WI low signal, isointensity or slightly high signal intensity T2WI, STIR fat suppression all showed obviously high signal, in which 20 patients with torn meniscus, collateral ligament injuries in 14 cases, anterior cruciate ligament injuries in 8 cases. The X-ray and CT examination of all the patients revealed no signs of fracture. The clinical symptoms and signs of all the patients significantly reduced or disappeared after treatment. Among 29 patients who were followed up from 1 to 12 months, 21 patients had abnormal MRI signal disappearing and 8 patients had abnormal signal weakened.

CONCLUSIONMRI can reveal pathological changes of bone contusion on the knee, and accurately determine ligaments around the joint and soft tissues injuries. Conventional MRI examination and fat-suppression sequence is the most valuable method of bone contusion.

Adolescent ; Adult ; Contusions ; diagnosis ; Diagnosis, Differential ; Female ; Humans ; Knee Injuries ; diagnosis ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

pHA2 plasmid sequence,with Bacterial Artificial Chromosome(BAC) vector and the GFP expression cassette, was introduced into the UL23(TK) gene of Pseudorabies virus(PRV)strain ZJ by homologous recombination,and the recombinant PRV (rPRV-HA2) was confirmed and isolated by plaque purification. The circular genome of rPRV-HA2 was electroporated into Escherichia coli strain DH10B and then the PRV BAC (pPRV) was recovered. The transfection of pPRV into VeroE6 cells resulted in productive infection. The rescued virus isolated following transfection was indistinguishable from rPRV-HA2 in cytopathic effects (CPE) and replication curve in vitro. The growth kinetics of the viruses indicated that partial deletion of TK gene and BAC vector insertion had no effect on the viral titre and plaque size in vitro. The PRV BAC system will enable quick and reliable manipulation of the viral genome for the functional investigation on the PRV genes and the development of PRV vector in vaccine.
Animals ; Cercopithecus aethiops ; Chromosomes, Artificial, Bacterial ; genetics ; Genome, Viral ; Herpesvirus 1, Suid ; genetics ; physiology ; Pseudorabies ; virology ; Recombination, Genetic ; Swine ; Swine Diseases ; virology ; Vero Cells ; Virus Replication

OBJECTIVETo explore the value of contrast-enhanced ultrasound (CEUS) in the diagnosis of thyroid occupied lesions with injection of sulphur hexafluoride microbubbles.

METHODSFifty nine cases of conventional ultrasonic diagnosis of thyroid lesions in patients with 73 lesions re-sulfur hexafluoride microbubble ultrasound contrast real-time inspection, the use of CEUS and contrast pulse sequencing (CPS).

RESULTSSeventy-three lesions were satisfied with the dynamic contrast perfusion imaging. Ultrasound contrast prompted the 15 lesions (13 cases) of malignant lesions by postoperative pathology confirmed papillary thyroid carcinoma, ultrasound images showed a low or mixed echo, boundary ambiguity, accompanied by Microcalcification foci, blood flow distribution I rank or grade III. Ultrasound prompted 58 lesions (46 cases) of benign lesions, blood flow distribution of grade II or III, the performance of a variety of two-dimensional ultrasound image, showing cystic, solid or liquid-solid mass, border clearance. Contrast with the surrounding thyroid tissue, lesionsor=20 mm papillary thyroid carcinoma, manifested prior to the beginning of the adjacent thyroid substance to enhance and dissection, showing the form of high-enhanced perfusion; thyroid follicular adenoma as early as the beginning of the adjacent thyroid substance to enhance and dissection, mass retained within the contrast agent longer time, showing the form of high-enhanced perfusion; nodular goiter enhanced for thyroid nodules and adjacent synchronization started to pick up in real terms, after the 6 lesions showed nodular contrast slightly earlier than the adjacent thyroid substance started to pick up, 28 lesions with synchronous real beginning of the adjacent thyroid dissection, 11 lesions of nodular dissipated early on (started receding time < or = 25 s) adjacent to thyroid substance.

CONCLUSIONThe real-time CEUS with injecting sulphur hexafluoride microbubbles is valuable in the diagnosis and differential diagnosis of the thyroid occupied lesions.

Adult ; Aged ; Contrast Media ; Female ; Humans ; Male ; Microbubbles ; Middle Aged ; Sulfur Hexafluoride ; Thyroid Neoplasms ; diagnostic imaging ; Ultrasonography ; Young Adult

OBJECTIVESTo study the relationship between the expression level of DLC-1 mRNA (located in 8p) and the invasion/metastasis of human hepatocellular carcinoma (HCC).

METHODSFifty-one surgical specimens of human HCC were divided into high-invasive and low invasive groups according to their clinicopathological features. DLC-1 mRNA expression was studied in the 51 HCC specimens as well as 5 different metastasis potential cell lines using real-time quantitative PCR (RQ-PCR).

RESULTSThe expression level of DLC-1 mRNA in HCC specimens with high invasiveness was significantly lower than that with low invasiveness (P < 0.05). The expression levels of DLC-1 mRNA were significantly different between non-metastatic (Hep3B and HepG2) and metastatic (MHCC97-H, MHCC97-L and HCCLM3) cell lines (P < 0.05). From MHCC97-L to HCCLM3, with an increase of invasiveness and metastatic potentials, the expression level of DLC-1 decreased correspondingly, and its expression level in HCCLM3 was significantly lower than that in MHCC97-L (P < 0.01).

CONCLUSIONThe expression of DLC-1 mRNA may play an important role in inhibiting the invasiveness and metastasis of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; GTPase-Activating Proteins ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mutagenesis, Site-Directed ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo study the relaxants effects of six extractions from Chinese Herbs (neferine, tetrandrine, kakonein, scutellarin, ginsenoside Rgl and ginsenoside Rb1) on the corpus cavernosum tissue of rabbit in vitro.

METHODSIsolated stripes of rabbit corpus cavernosum tissue were precontracted with 10(-5) mol/L phenylephrine (PE). Relaxation in response to cumulative doses of six extracts at (10(-8) - 10(-3)) mol/L was determined.

RESULTSOn rabbit cavernosal muscle stripes precontracted with PE, neferine, tetrandrine, kakonein and scutellarin showed dose dependent relaxation. IC50 values were 4.60 x 10(-6), 3.73 x 10(-5), 8.03 x 10(-4) and 3.33 x 10(-3) mol/L, respectively. However, in the meantime, it was found that the relaxant effects of ginsenoside Rgl and ginsenoside Rbl less significant to stripes precontracted with PE. When the final concentration was 10(-3) mol/L, the relaxations were only (16.32 +/- 5.45)% and (11.21 +/- 3.10)%.

CONCLUSIONAmong the six extracts which showed relaxant effects to rabbit cavernosal muscle stripes precontracted with PE, neferine had greater functions than the other five extracts.

Animals ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; In Vitro Techniques ; Male ; Muscle Contraction ; drug effects ; Penis ; drug effects ; physiology ; Rabbits

OBJECTIVETo evaluate the effects of red clover isoflavones on the proliferation and apoptosis of human benign prostatic hyperplasia (BPH) stromal cells.

METHODSWe treated human prostate stromal cells with red clover isoflavones at the concentration of 12.5, 25, 50 and 100 microg/ml, and established a PBS blank control, a dimethyl sulphoxide (DMSO) negative control and four finasteride positive control groups (at the concentration of 12.5, 25.0, 50.0 and 100.0 microg/ml). We determined the effects of different concentrations of red clover isoflavones on the proliferation of the cells by MTT assay and on their apoptosis by Annexin V/PI double staining flow cytometry.

RESULTSRed clover isoflavones inhibited the proliferation of the BPH stromal cells by 18.86% at 25.0 microg/ml, compared with 5.17% in the blank control group (P < 0.05), and more obviously at a higher concentration. At 50.0 microg/ml, red clover isoflavones exhibited a weaker inhibitory effect than finasteride (28% vs 69.88% , P < 0.05). Annexin V/PI double staining flow cytometry showed that red clover isoflavones at 25.0 microg/ml induced the apoptosis of the prostate stromal cells by (18.54 +/- 2.5)%, with significant differences from the negative control and blank control (P < 0.01).

CONCLUSIONRed clover isoflavones can inhibit the proliferation and promote the apoptosis of human BPH stromal cells.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Humans ; Isoflavones ; pharmacology ; therapeutic use ; Male ; Plant Extracts ; pharmacology ; therapeutic use ; Prostate ; cytology ; drug effects ; Prostatic Hyperplasia ; drug therapy ; pathology ; Stromal Cells ; drug effects ; Trifolium ; chemistry