A PCR method was used to amplify the sequence encoding the mature peptide of?-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of?-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.

Trehalose is a naturally occurring non-reducing disaccharide. This sugar has been known as a protector of biomembrane and macromolecules. And it has been widely used in foods, medicines, cosmetics. Trehalose synthase (TreS) is an intramolecular transglycosylase. It can catalyze the conversion of maltose into trehalose in one step. As the simple protocol and cheap substrate, it is very prospect in the industrial production. Here we have discussed the properties, catalytic mechanism and gene-engineering of TreS, and also proposed some solutions to its present research problems.

Animals ; Barbiturates ; Coloring Agents ; Fallopian Tubes ; cytology ; embryology ; physiology ; Female ; Isoxazoles ; Male ; Membrane Potentials ; Mice ; Mice, Inbred Strains

Objective To investigate the clinical features and distribution of G types of rotavirus diarrhea in infants and children in Suzhou and Maanshan areas.Methods 1267 stool specimens were collected from children under 5 years with acute diarrhea. Enzyme-linked immunosorbant assay (ELISA) and nested polymeras chain reaction (Nested-PCR) were used to detect rotavirus and serotypes.Results In 1267 stool specimens, 378 were detected RV positive, and serotype G was the prevalent type in the two cities with number of 250 and the G3 and G1 types were the most prevalent which accounted for 40%(100/250) and 44.40 % (111/250), respectively. While the G2 ,G4, G9 types and mixed infections were 8,3,3,4 and the un-identified type was 8.40%(21/250). G1 type was the most prevalent in Maanshan are which accounted for 58.54% in all cases,while the G3 type accounted for the most of case(47.85%) in Suzhou area.Conclusions The results indicated that rotavirus was the most important etiologic agent in Maanshan and Suzhou areas,G1 and G3 were the most prevalent types,respectively. J Appl Clin Pediatr,2005,20(3):208-210

BACKGROUND/AIMS: To evaluate the enhancement patterns of liver metastases and their influencing factors using dynamic contrast-enhanced ultrasound (CEUS). METHODS: A total of 240 patients (139 male and 101 female; 58.5±11.2 years of age) diagnosed with liver metastases in our hospital were enrolled in this study to evaluate tumor characteristics using CEUS. A comparison of enhancement patterns with tumor size and primary tumor type was performed using the chi-square test. The differences between quantitative variables were evaluated with the independent-sample t-test and one-way analysis of variance. RESULTS: The enhancement patterns of liver metastases on CEUS were categorized as diffuse homogeneous hyperenhancement (133/240, 55.4%), rim-like hyperenhancement (80/240, 33.3%), heterogeneous hyperenhancement (10/240, 4.2%), and isoenhancement (17/240, 7.1%). There were significant differences in the enhancement patterns during the arterial phase based on the nodule size (p=0.001). A total of 231 of the nodules showed complete washout during the portal phase, and 237 nodules were hypoenhanced during the delayed phase. The washout time was correlated with tumor vascularity, with a longer washout time observed in hypervascular metastases compared to hypovascular metastases (p=0.033). CONCLUSIONS: Diffuse homogeneous hyperenhancement followed by rapid washout was the most common enhancement pattern of liver metastases on CEUS and was affected by the nodule size and tumor vascularity. Small metastases were prone to show diffuse homogeneous hyperenhancement. Hyper-vascular metastases showed a significantly longer washout time compared to hypovascular metastases.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Contrast Media/*therapeutic use ; Female ; Humans ; Liver/diagnostic imaging/pathology ; Liver Neoplasms/blood supply/*diagnostic imaging/secondary ; Male ; Middle Aged ; Neovascularization, Pathologic/diagnostic imaging ; Ultrasonography/*methods ; Young Adult

OBJECTIVETo study the expression changes of neuroglobin in rats with the model of diffuse traumatic brain injury and explore the relationship between the neuroglobin and neuron apoptosis in traumatic brain injury.

METHODSThe diffuse traumatic brain injury of rats was induced by the Marmarou's 'weight-drop' device. And the immunohistochemical technique was used to detect the expression changes of neuroglobin and neuron apoptosis in rat brain at different time points post-injury.

RESULTSThe expression of neuroglobin increased twice and reached peaks at 2 hours and 72 hours post-injury respectively. And the increased expression of neuroglobin from 30 minutes to 1 hour post-injury and from 48 hours to 72 hours post-injury accompanied with the decreased expression ratio of Bax to Bcl-2.

CONCLUSIONThe increased expression of neuroglobin in traumatic brain injury informed us that neuroglobin had anti-apoptosis action in post-injury neuron. It could protect the neuron from traumatic stress and secondary ischemia and hypoxia insults during ultra-early and acute stages.

Animals ; Apoptosis ; physiology ; Brain ; metabolism ; pathology ; Brain Injuries ; metabolism ; pathology ; Globins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Neurons ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective: To construct and identify lentiviral vector containing human ILK-shRNA and mda7 gene.Methods:Based on the human ILK gene sequences , RNAi target sequences were designed and cloned into the lentiviral vector pSicoR -eGFP by restriction endonuclease HpaⅠand XhoⅠdouble digestion and T 4 DNA ligase ligation . Based on the human mda7 gene sequences , PCR primers were designed to clone the full-length mda7, and were cloned into the lentiviral vector pLVX-Puro.After the candidate clones were identified by DNA sequencing , the recombinant plasmid and the three packaging plasmids were co-transfected into the human embryonic kidney 293T cells by lipofectamine 2000 to produce the lentiviral particles .Human prostate cancer PC-3 cells were infected with the constructed lentiviral vector .The ILK and mda7 expression levels in PC-3 cells were quantified by qPCR and Western blot , respectively .The effect of ILK and mda7 on proliferation and migration of PC-3 cells were assessed by MTT method and Transwell assay, respectively.Results: ILK-pSicoR-eGFP and mda7-pLVX-Puro lentiviral vectors were successfully constructed .Strong green fluorescence was observed in the 293T cells under the fluorescent microscope after co-transfection of 293T cells with 4 plasmids of lentiviral vector .The transfection efficiency of the collected virus exceeded 90%in the 293T cells and the PC-3 cells were infected with the lentiviral particles with high efficiency .The A and B lentiviral vector inhibited the expression of ILK at both the mRNA and protein levels in PC-3 cells significantly .The mda7-pLVX-Puro lentiviral vector increased the expression of mda 7 in PC-3 cells, and the ability was maintained for one month.Within 96 h, ILK and mad7 significantly inhibited the proliferation and migration of PC-3 cells ( Ps <0.05 ) . Conclusion: The lentiviral vectors of ILK knockdown and mda 7 over-expression have been successfully constructed and identified . The recombinant lentivirus can efficiently infect human prostate cancer PC -3 cells, in which ILK expression is inhibited and mda 7 is over-expressed .

OBJECTIVETo investigate the value of real-time harmonic contrast-enhanced ultrasound (CEUS) in the diagnosis of atypical liver abscesses.

METHODSWe used Technos DU8 system with CEUS technique to examine 32 patients with suspected liver masses. Mechanical index was set at 0.08- 0.11. The contrast agent SonoVue was injected as a bolus (2.4 ml, < 5s) in the antecubital vein.

RESULTSA total of 36 lesions were studied in 32 patients, including 30 patients with single nodule and 2 patients with multiple nodules. All lesions with liver abscess were confirmed by biopsy. The whole-enhanced pattern in all lesions was clearly displayed in the arterial phase after the administration of SonoVue. The mean time of the beginning enhancement was (14.48 +/- 3.62) s, the mean peak enhancement time was (22.92 +/- 4.35) s, and the mean time of hypoechoic appearance of lesion was (80.30 +/- 35.30) s. The lesions with hypoechoic pattern (37.5%, 12/32) and isoechoic pattern (62.5%, 20/32) in the portal phase were detected by CEUS. The characteristic sign with honeycomb pattern on CEUS was shown in 30 (93.8%) liver abscess lesions.

CONCLUSIONCEUS is valuable in the diagnosis of atypical liver abscesses.

Contrast Media ; Humans ; Liver Abscess ; diagnostic imaging ; Ultrasonography

OBJECTIVETo review the preliminary clinical experience with high-field-strength intra-operative magnetic resonance imaging (iMRI) in the endoscopic chordoma operation with transsphenoidal or transoral approach.

METHODSFrom January 2009 to December 2010, 23 patients [range, 29 - 64 years, mean age (42 ± 3) years] of chordoma were operated with endoscopic transsphenoidal or transoral approach and examined intraoperatively with a movable 1.5 T iMRI magnet. Tumor size range was 2.0 - 5.7 cm, mean (3.5 ± 0.8) cm. A navigation system based on iMRI was used in 20 cases.

RESULTSiMRI scan were performed in each operation from 1 time to 5 times. Neuronavigation system were used in 20 operations and the data renewed in 12 cases by the information from iMRI. In 15 of 23 patients, iMRI had revealed residual lesions and resulted in 12 cases further treatment, eventually, 9 tumors were totally removed and 3 tumors were further removed. The ratio of total removal tumor was enhanced to 73.9% (17/23) from 34.8% (8/23). Among 15 cases of partial chordoma removal detected by scanning in operation, 9 were huge chordoma. The residual of huge chordoma detected by scanning in operation was 9/11, and other chordoma contributed to 6/12. There were no iMRI related safety issue or accident recorded in this study.

CONCLUSIONSHigh-field-strength iMRI provide high-quality images of tumor resection that allows intraoperative modification of the surgical strategy. Combined with the navigation system, iMRI is helpful to maximize the resection of the chordoma and benefit for the safety of endoscopic operation.

Adult ; Chordoma ; surgery ; Endoscopy ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neuronavigation ; methods ; Pituitary Neoplasms ; surgery ; Sphenoid Sinus ; surgery

OBJECTIVETo investigate the prevalence and epidemiologic characteristics of seronegative spondyloarthritis (SpAs) in Chinese army force in different areas.

METHODS4-phase survey was conducted in 21 750 Chinese army, including: face-to-face interviews with standardized COPCORD questionnaires (Phase I screening); further examination on the suspected cases; identification of inflammatory joint and spinal diseases (Phase II); identification of SpAs (AS and uSpA) by more than two experienced specialists in rheumatology; further examination with X-rays and laboratory detection of HLA-B27 (Phase III); and data analysis (Phase IV).

RESULTSAmong 21 750 army men, 21 cases of RA, 106 cases of SpAs were identified, with prevalence rates of 0.966 per thousand, 4.87 per thousand respectively. In 106 cases of SpAs, there were 46 cases of ankylosing spondylitis (AS), 52 cases of undifferentiated SpAs (uSpAs) with the prevalence rates of 2.11 per thousand and 2.39 per thousand respectively. Few cases of reactive arthritis (ReA) and Reiter's syndrome (RS) were identified (6 and 1 cases respectively). The prevalence of AS, uSpAs were higher in navy than that in the ground force or the air force. Soldiers in cold and damp areas had higher prevalence rates than that in the plain and drought areas.

CONCLUSIONThe prevalence of SpA (especially AS and uSpA) in Chinese army force was similar to that in the civilians. SpA (AS and uSpA) was more prevalent seen in the Navy. The incidence of SpA (AS and uSpA) was influenced by environmental factors such as coldness and dampness.

Adult ; China ; epidemiology ; Humans ; Lumbar Vertebrae ; Male ; Military Personnel ; Prevalence ; Sacrum ; Spondylarthritis ; epidemiology ; etiology