A novel gene,located between dnrX and drrB in the genome of daunorubicin-producing strain Streptomyces coeruleorubidus SIPI-1482,was cloned and named as dauW.The full sequence of dauW was submitted to GenBank(Accession No.EF523565).Blast result indicated that it showed high homology with dnrW in GenBank.The exact function of dauW is as yet unknown despite the possibility that it might belong to a family of FAD-dependent oxidoreductases on the basis of conserved domain analysis.dauW was cloned into expression plasmids pET-28a(+)and pET-32a(+),respectively,and was successfully expressed in E.coli DE3 after induction with IPTG.The preliminary results of the expression of dauW suggested that it might be involved in the self resistance in Streptomyces coeruleorubidus due to the increased resistance to daunorubicin in the E.coli host.

TDP-4-ketohexulose reductase, encoded by dnmV, is important in daunorubicin biosynthesis. To obtain a daunorubicin block mutant, double cross-over plasmid pYG817 was constructed by inserting apramycin resistant gene and amplified dnmV together with upstream dnmU into vector pUC18. dnmV was successfully disrupted after transformation of daunorubicin-producing strain SIPI-1482 by pYG817. Daunorubicin was absent from metabolites of the resulting transformant, and its biosynthesis can be reconstituted by introducing dnmV expression plasmid into the disruptant, although the yield is lower than wild-type SIPI-1482, according to HPLC analysis. This mutant can be a good candidate for production of anthracycline such as epi-daunorubicin by introducing exogenous gene into the host.

To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.

TTI gene coding for Tsetse thrombin inhibitor was modified with E.coli bias codon and expressed in Escherichia coli with high efficiency.Recombinant protein was purified to more than 98% purity.Assay for enzyme activity determination was set up.The result showed that the fusion protein exhibited inhibiting activity for thrombin.Inhibitory rate of purified TTI was 73% when concentration of thrombin and substrate was 10U/ml and 250?mol/L respectively.Inhibition pattern was determined as competitive with Ki at 35?mol/L.

?-lactamase inhibitor research is popular for its potential on ?-lactam antibiotics resistant strain.A ?-lactamase binding peptide SIPIS04-01 was obtained by the yeast two-hybid system.In vitro assay showed that it can inhibit the ?-lactamase activity.In order to improve the expression level of the recombinant peptide,a two-copy expression plasmid pYG563 was constructed by random orientation tandem repeat method after codon modification,the two-copy plasmid was successfully expressed and the product was increased by 48.4% than that of one-copy plasmid.Purified peptide showed inhibitory activity against TEM-1 ?-lactamase in vitro and the inhhibitory constant Ki was measured.

OBJECTIVETo follow up the electrocardiographic and cardiac autonomic function changes after percutaneous transluminal septal myocardial ablation (PTSMA) in patients with hypertrophic obstructive cardiomyopathy (HOCM).

METHODSBaseline, 3 days and 3 years post procedure 12-lead electrocardiographic and 24-hour Holter electrocardiographic recordings including PR interval, QRS duration, cardiac conduct block, QT, QTd, QTcd, JT, JTd, JTcd, heart rate variability (HRV) data (SDNN, SDANN, HF, rMSSD, PNN50, LF, HF, LF/HF) were analyzed in 26 patients with HOCM receiving PTSMA.

RESULTThe PTSMA procedure was successful in all 26 patients. One patient developed complete atrioventricular block requiring permanent pacing. The PR interval was significantly prolonged 3 days after ablation and recovered 3 years post procedure. Right bundle branch block was seen in all patients 3 days after post procedure and in 24 patients at 3 years post procedure. The QRS duration was significantly prolonged at 3 days and 3 years post procedure. There was persistent QT interval prolongation up to 3 years and transient QTd, QTcd prolongation (prolonged at 3 days and returned to baseline at 3 years after ablation) while JT, JTd, JTcd were not significantly changed after PTSMA. LF, HF, rMSSD and PNN50 were significantly increased while LF/HF, SDNN, SDANN remained unchanged post procedure.

CONCLUSIONPTSMA is a safe and effective therapy option for HOCM. Right bundle branch block was the main electrocardiographic change post procedure and PTSMA could partly restore the heart sympathovagal balance by improving vagal activity.

Adult ; Autonomic Nervous System ; physiopathology ; Cardiomyopathy, Hypertrophic ; physiopathology ; therapy ; Catheter Ablation ; methods ; Electrocardiography ; Female ; Follow-Up Studies ; Heart Septum ; Humans ; Male ; Middle Aged

OBJECTIVETo explore the changes of metastatic potential of residual hepatocellular carcinoma (HCC) after in vivo chemotherapy and its mechanism.

METHODSNude mouse models of orthotopic HCC in the nude mouse livers was established using human hepatocellular carcinoma cell line MHCC97L cells. Oxaliplatin (10 mg/kg, once per week) was administered intraperitoneally (i.p.) to mice in the trial group. Mice in the control group received 0.2 ml of 0.9% sodium chloride on the same days. On day 7 after the third injection, all mice were sacrificed and tumor fragments of equal volume (2 mm×2 mm×2 mm) from each mouse of the oxaliplatin-treated and untreated groups were reinoculated into the livers of each new recipient mouse correspondingly. The growth, metastasis and molecular phenotype of the reinoculated tumors in both groups were determined.

RESULTSIn the new recipient mice, compared with untreated tumors, oxaliplatin pre-treated tumors grew significantly slower [(2624.59 ± 491.60) mm(3) vs. (3849.72 ± 827.09) mm(3), P < 0.001], but gave more spontaneous metastasis to the lung (10/12 vs. 3/12, P = 0.012). A decreased expression of E-cadherin and increased expression of N-cadherin, vimentin and transcription factor Snail were detected in the oxaliplatin pre-treated tumors by immunohistochemistry, which provided the evidence of epithelial mesenchymal transition (EMT) in these tumors.

CONCLUSIONResidual hepatocellular carcinomas after in vivo chemotherapy grow slower but gain enhanced metastatic potential to the lung, associated with epithelial mesenchymal transition.

Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Cadherins ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; secondary ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Humans ; Liver Neoplasms ; drug therapy ; metabolism ; pathology ; Lung Neoplasms ; drug therapy ; secondary ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Neoplasm, Residual ; drug therapy ; metabolism ; secondary ; Organoplatinum Compounds ; therapeutic use ; Snail Family Transcription Factors ; Transcription Factors ; metabolism ; Tumor Burden ; Vimentin ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo evaluate the effect of glutamine granules on protein metabolism in severe burns and trauma patients.

METHODS120 patients with severe burns, multiple trauma and post operation who met the requirements of the protocol joined this double-blind randomized controlled, multi-center clinical trail. Patients were randomly divided into two groups: placebo control group (P group, 60 patients) and glutamine granules treatment group (GLN group, 60 patients). There was isonitrogenous and isocaloric intake in both groups, GLN and P group patents had been given glutamine granules or placebo (glycine) at 0.5 g.kg(-1).d(-1) for 7 days, respectively. The level of plasma glutamine, protein and urine nitrogen exclude were determined, wound healing rate of burn area and hospital stay were recorded, and then observed the complication and side effect.

RESULTSAfter 7 days of taking glutamine granules orally, plasma GLN concentration was significant higher than that in P group (592.50 +/- 185.23 micro mol/L vs. 407.41 +/- 190.22 micro mol/L) (P < 0.01). Plasma prealbumin and transferrin in GLN group were significant higher than those in P group (P < 0.01), but the concentration of total protein and albumin were no marked changes compare with P group (P > 0.05). The capacity of urine nitrogen exclude in GLN group were significant lower than that in P group. Additional, the wound healing rate was faster and hospital stay days was shorter than P group (P < 0.05), and the occurrence of glutamine granules side effect was seldom.

CONCLUSIONTaking glutamine could promote protein synthesis, abate protein decompose, ameliorate wound healing rate and reduce hospital stay obviously.

Adult ; Burns ; drug therapy ; metabolism ; Double-Blind Method ; Female ; Glutamine ; blood ; therapeutic use ; Humans ; Length of Stay ; Male ; Middle Aged ; Nitrogen ; urine ; Proteins ; metabolism ; Treatment Outcome ; Wounds and Injuries ; drug therapy ; metabolism

OBJECTIVETo evaluate the effect of glutamine granules on immunofunction in severe burns and trauma patients.

METHODSOne hundred and twenty patients with severe burns, multiple trauma and post operation who met the requirements of the protocol joined this double-blind randomized controlled, multi-center clinical trail. Patients were randomly divided into two groups: placebo control group (P group, 60 patients) and glutamine granules treatment group (GLN group, 60 patients). There was isonitrogenous and isocaloric intake in both groups. GLN and P group patients had been given glutamine granules or placebo (glycine) at 0.5 g.kg(-1).d(-1) for 7 days, respectively. The level of plasma glutamine and some index of immunofunction were determined, and the complication and side effect were also observed.

RESULTSAfter 7 days of taking glutamine granules orally, plasma GLN concentration was significantly higher than that in P group [(593 +/- 185) micromol/L vs (407 +/- 190) micromol/L)] (P < 0.01). IL-2 level, CD(4)/CD(8) ratio, PMN swallow ratio in GLN group were significantly higher than those in P group (P < 0.05-0.01), but the concentration of IgG, IgM, C(3)/C(4) were not significantly different when compared with P group (P > 0.05). In addition, the occurrence of side effect in both groups was seldom.

CONCLUSIONTaking glutamine granules could increase plasma GLN concentration, enhance body immunofunction, and using glutamine granules is safe.

Administration, Oral ; Adolescent ; Adult ; Double-Blind Method ; Female ; Glutamine ; adverse effects ; blood ; therapeutic use ; Humans ; Male ; Middle Aged ; Wounds and Injuries ; blood ; drug therapy ; immunology

Female ; Gallstones ; pathology ; surgery ; Humans ; Hyperparathyroidism, Primary ; diagnosis ; surgery ; Pancreatitis ; pathology ; surgery ; Young Adult