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

Objective To observe the clinical efficacy and safety of deprogesterone tablets in the treatment of climacteric syndrome.Methods A total of 86 women with climacteric syndrome were randomly divided into control group and treatment group with 43 cases per group.Control group received placebo 10 mg per time,qd,orally.Treatment group was treated with deprogesterone 10 per time,qd,orally.Two groups were treated for one month.The clinical efficacy,the levels of sex hormone and blood lipid,adverse drug reactions were compared between two groups.Results After treatment,the total effective rates of treatment and control groups were 93.02％ (40 cases/43 cases) and 72.09％ (31 cases/43 cases) with statistically significant difference (P ＜ 0.05).After treatment,the main indexes in treatment and control groups were compared:luteinizing hormone were (11.70 ± 1.13) and (28.36 ± 4.48) U · L-1,follicle stimulating hormone were (36.31 ±4.24) and (60.77 ± 7.75) U · L-1,estradiol were (71.76±9.37) and (45.43 ±5.21)pmol · L-1,triglyceride were (1.23 ± 0.24) and (1.72 ± 0.20) mmol · L-1,total cholesterol were (4.94 ± 0.66) and (5.82 ± 0.58) mmol · L-1,low density lipoprotein cholesterol were (2.64 ± 0.36) and (3.25 ±0.31)mmol · L-1,high density lipoprotein cholesterol were (1.66 ±0.21) and (1.38 ±0.13) mmol · L-1,the differences were statistically significant (all P ＜ 0.05).The adverse drug reactions in treatment group were based on insomnia (2 cases) and headache (2 cases).And there was no adverse drug reaction in the control group.The incidences of adverse drug reactions in treatment and control groups were 9.30％ and 0 without significant difference (P ＞ 0.05).Conclusion Deprogesterone tablets have a definitive clinical efficacy in the treatment of climacteric syndrome,which can effectively adjust the levels of sex hormone and blood lipid,without increasing the incidence of adverse drug reactions.

OBJECTIVE To isolate and purify an active polysaccharide of Momordica charantia L.and investigate its struc-tural characterization and immunomodulatory and antitumor activities.METHODS A polysaccharides was isolated from Mo-mordica charantia L.in hot water extraction followed by dialysis and purification through anion exchange cellulose chromatog-raphy and gel filtration chromatography,and its molecular weight was measured by high performance gel permeation chroma-tography.Additionally,Fourier transform infrared spectroscopy(FT-IR)and nuclear magnetic resonance(NMR)were used to determine its partial structural characterization,and its immunomodulatory and antitumor activities were measured by cell and molecular biological techniques and pharmacological methods.RESULTS A heteropolysaccharide named MCP-2 with molecu-lar weight of 745 kDa was isolated and purified,which was shown to be consisted of rhamnose,galacturonic acid,galactose, xylose,and arabinose with corresponding molar ratio being 1.1 9 ∶ 1 5.97 ∶ 3.40 ∶ 0.73 ∶ 0.94.Furthermore,MCP-2 was found to promote the lymphocyte proliferation of normal mice and to raise the levels of NO,IL-1β and TNF-α produced by macrophage RAW264.7.Intriguingly,MCP-2 presented the antitumor activity against S180 cells in vivo as well.CONCLU-SION All experimental data showed that MCP-2 had apparent immunologic enhancement and antitumor activities.

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

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 observe the influence of high blood glucose fluctuation on the endothelial function of type 2 diabetes mellitus (T2DM) rats and the effects of Panax Quinquefolius Saponin (PQS) of stem and leaf.

METHODSThe T2DM model was induced by intraperitoneal injection of a small dose of streptozotocin (STZ, 35 mg/kg) plus high fat and high caloric laboratory chow. Then, diabetic rats were divided into steady high blood glucose (SHG) group and fluctuant high blood glucose (FHG) group according to fasting blood glucose coefficient of variation (FBG-CV), and then, the FHG group rats were divided into 4 groups according to the level of FBG-CV and fasting blood glucose: PQS 30 mg/(kg·d) group, PQS 60 mg/(kg·d) group, metformin hydrochloride control (MHC) group, and FHG control group, 10 in each group. Meanwhile, 10 rats without any treatment were used as normal control (NOR) group. Eight weeks later, the aortic arteries histology, plasma hepatocyte growth factor (HGF), and serum nitric oxide (NO), endothelin-1 (ET-1), tumor necrosis factor α (TNF-α), and soluble intercellular adhesion molecule 1 (sICAM-1) were measured.

RESULTSIn comparison with the NOR group, the level of plasma HGF and serum NO, ET-1 and TNF-α, and sICAM-1 in SHG and FHG control groups were all significantly increased (P<0.01); in comparison with the SHG group, plasma HGF and serum NO, ET-1, TNF-α, and sICAM-1 in FHG group were all significantly increased further (P<0.01 or P<0.05); meanwhile, in comparison with the FHG control group, the level of plasma HGF and serum NO, ET-1, TNF-α, and sICAM-1 in PQS and MHC groups were all decreased significantly (P<0.01). However, comparison of the aortic arteries histology among groups showed no significant differences either before or after treatment.

CONCLUSIONBlood glucose fluctuation could facilitate the development of vascular endothelial dysfunction in T2DM rats, while PQS could improve the endothelial function of T2DM rats with high blood glucose fluctuation, which may be related to its effects of relieving vessel stress, decreasing vasoconstrictor ET-1 production, preventing compensated increase of NO, and reducing inflammatory reaction.

Animals ; Aorta ; drug effects ; pathology ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Diabetes Mellitus, Type 2 ; blood ; drug therapy ; Endothelin-1 ; blood ; Endothelium, Vascular ; drug effects ; physiopathology ; Hepatocyte Growth Factor ; blood ; Intercellular Adhesion Molecule-1 ; blood ; Male ; Nitric Oxide ; blood ; Panax ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rats ; Saponins ; pharmacology ; therapeutic use ; Solubility ; Tumor Necrosis Factor-alpha ; blood