One alkaline protease from Actinomucor elegans AS3.2778 was purified protein. The enzyme was purified using ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and size exclusion chromatography method, and its properties were also investigated. The molecular weight of this enzyme is 32 kDa with SDS-PAGE method, optimum temperature is 60℃, optimum pH is 8.5 to 10.5, it is stable in the pH range of 6.0 to 9.0 at < 40℃ temperature, and being completely inhibited by the serine protease inhibitor, PMSF, indicated that it belongs to the serine protease family. Specificity test indicated this protease has extensive selectivity to peptide bones, especially to peptide bones composed of Leucine residue.

Objective To investigate the effect of internal fixation with proximal femoral nail antirotation (PFNA) and bone cement on peritrochanteric tumors due to metastatic carcinoma.Methods Clinical data of 19 patients with peritrochanteric tumors due to metastatic carcinoma during June 2007 and July 2013 treated with PFNA and bone cement were retrospectively analyzed.Results Visual analogue scale(VAS) was (8.37 ± 1.12) scores before surgery,and (2.58 ± 1.26) scores after 3 d surgery.There was significant difference (t =22.45,P ＜ 0.05).Conclusion For patients with peritrochanteric tumors due to metastatic carcinoma,internal fixation with PFNA and bone cement is an effective way to relieve pain,restore and improve hmb function and enhance quality of life.

Objective To detect the expression level of Taurine up?regulated gene 1( TUG1) in the re?nal cell carcinoma and paired paracancerous normal tissues,then explore the relationships between the expression level of TUG1 and clinical characteristics.Methods RNA was Extacted from the resected renal cell carcinoma tissues and paired paracancerous normal tissues of 46 patients respectively,by reverse transcription to get cDNA, the expression level of the TUG1 was detected by RT?qPCR, the relationship between the expression level of TUG1 and the clinicopathological characteristics was analyzed by statistically software. Results The expression of TUG1 in renal cell carcinoma was obviously lower than that in paired paracancerous normal tissues(0.533±0. 027 vs. 1.000±0.298,t=-3.350,P<0.01).The△CT value of Tug1 in 46 cases of renal cell carcinoma after log?arithmic transformation,the minimum value was -5.535,maximum was 3.085,average value was -0.908,with the average of -0.908 as a dividing line,46 cases of renal cell carcinoma with 25 cases (54.34%) were down regulated the expression.The expression level of TUG1 of patients with renal carcinoma have no significant corre?lation with age,sex,type of renal cell carcinoma,TNM staging and UICC/AJCC staging(P>0.05).Conclusion The expression of TUG1 in renal cell carcinoma tissues are down?regulated,which also suggest that it may be re?lated to the tumorigenesis and development of renal cell carcinoma.

Child, Preschool ; Chromosomes, Human, Pair 22 ; Chromosomes, Human, Pair 8 ; Chromosomes, Human, Pair 9 ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Translocation, Genetic

Objective To explore the effects and safety of hemoperfusion(HP) therapy on tetramine poisoning in infants.Methods Thirty-five infants with tetramine poisoning were divided into two groups: HP group(n=18) and non HP group(n=17).The changes of blood tetramine concentration and clinical symptom improving of both groups after the treatment were observed together with the adverse effects of HP group.Results The average blood tetramine concentration of HP group was higher than that of non HP group(342.2?333.4 vs 117.9?50.8 ?g/L,P

0.05),while the level of IL-8,TNF-? and endotoxin changed significantly during CRRT(Pa

OBJECTIVETo determine the frequencies and significance of myeloid-derived suppressor cells (MDSCs) and T-helper 17 (Th17) cells in peripheral blood of young children with recurrent wheezing.

METHODSThirty young children with an acute exacerbation of recurrent wheezing were randomly enrolled. Twenty age-matched children with bronchopneumonia (pneumonia group) and 23 age-matched preoperative children with non-infectious or non-neoplastic diseases (hernia or renal calculus) (control group) were selected. The frequencies of MDSCs and Th17 cells in the peripheral blood were measured using flow cytometry and their correlation was determined by the Spearman's correlation coefficient.

RESULTSThe percentage of MDSCs in nucleated cells was significantly higher in the wheezing group than in the pneumonia and control groups (P<0.05), and it was significantly higher in the pneumonia group than in the control group (P<0.05). The percentage of Th17 cells in mononuclear cells was significantly higher in the wheezing group than in the pneumonia and control groups (P<0.05), but it showed no significant difference between the pneumonia and control groups (P>0.05). The frequency of MDSCs was positively correlated with the frequency of Th17 cells in the wheezing group (r=0.645, P<0.01).

CONCLUSIONSMDSCs and Th17 cells may contribute to the pathogenesis of recurrent wheezing in young children.

Child, Preschool ; Female ; Humans ; Infant ; Leukocytes, Mononuclear ; immunology ; Male ; Myeloid Cells ; immunology ; Recurrence ; Respiratory Sounds ; immunology ; Th17 Cells ; immunology

To explore the value of detection of PML/RARalpha gene rearrangement on bone marrow smears (BMS) by dual-color fluorescence in situ hybridization (D-FISH) for the diagnosis of acute promyelocytic leukemia (APL), the locus-specific probes for PML and RARalpha genes labeled directly and respectively by Spectrum Green and Spectrum Orange and the D-FISH technique were used to detect the PML/RARalpha gene rearrangement on BMS in 27 suspected APL patients. The results were compared with that of conventional cytogentics (CCG) and reverse transcriptase polymerase chain reaction (RT-PCR). The results showed that out of 18 newly diagnosed patients 14 were found having t(15;17) translocation by CCG and PML/RARalpha gene rearrangement were confirmed by BMS-D-FISH and RT-PCR. Thus, their APL diagnosis was determined; out of 4 patients in whom t(15;17) translocation was not detected by CCG, one had positive BMS-D-FISH and RT-PCR results, thus, this case was considered as having a cryptic t(15;17) translocation, three had negative BMS-D-FISH and RT-PCR results, thus, they were diagnosed as having acute myeloid leukemia rather than APL. In 9 cases with remission, one case with partial remission was found having t(15;17) translocation by CCG, and he had positive BMS-D-FISH and RT-PCR results, the other 8 patients (6 cases with normal karyotype and 2 cases without CCG examination) displayed different BMS-D-FISH and RT-PCR results: negative in 6 cases, but positive in 2 cases. The 2 cases were believed that they survived with minimal residual disease (MRD). It is concluded that BMS-D-FISH is a sensitive and reliable method for the detection of PML/RARalpha rearrangement. It is helpful for diagnosing APL and monitoring its MRD, and especially fit to those patients presenting a cryptic translocation or with failed cytogenetics, lacking suitable material for RT-PCR, as well as needing retrospective study.
Adolescent ; Adult ; Bone Marrow Cells ; metabolism ; Female ; Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Leukemia, Promyelocytic, Acute ; diagnosis ; genetics ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Receptors, Retinoic Acid ; genetics ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity

OBJECTIVETo use a new kind of fixing material, i.e. Sol-Gel organic-inorganic hybridized material to immobilize bacterium to detect Biochemical oxygen demand quickly.

METHODSThe biosensor was fabricated using a thin film in which Hansenula anomala was immobilized by sol-gel and an oxygen electrode. The optimum measurement for biochemical oxygen demand was at pH 7.0; 28 degrees C; response time 3 - 12 min. Pure organic compound, sewage and rate of recovery were detected with the biosensor.

RESULTSIt shows that the BOD biosensor can be used to detect many organic compounds such as amino acid, glucide. It is suitable to monitor sewage and industrial waste water which has low level alcohols and phenols. The microbial membrane can work 3 months and remain its 70% activity. It is measured that the rate of recovery of BOD is between 90% to 105% in sewage.

CONCLUSIONThe study confirmed the effectiveness and usefulness of BOD sensor, which is quick, convenient, low cost and reliable with little interference.

Bacteria ; Biosensing Techniques ; instrumentation ; Cells, Immobilized ; Gels ; Membranes, Artificial ; Nylons ; Oxygen ; analysis ; Sewage ; analysis ; microbiology

AIMTo reconstruct of a tissue engineering skin in vitro for the study of the use of drug percutaneous penetration and metabolism.

METHODSDermal fibroblasts were embedded in collagen type I. HaCaT cells were seeded on the top of the gel. The skin was generated through air-liquid interface culture. Effects of various culture media on tissues morphology were investigated. Sections of the cultured skin were stained with hematoxylin and eosin and examined under microscope. Permeation and metabolism of ketoprofen and its isopropyl ester through the cultured skin were investigated.

RESULTSHaCaT cells initially developed a multilayer epithelium at the air-liquid interface, but it showed a parakeratotic stratum corneum. Vitamin C enhanced cell proliferation obviously. Vitamin D3 promoted cell differentiation. And estradiol showed little effect on the tissue engineering skin. Ketoprofen isopropyl ester was hydrolyzed into ketoprofen when penetrated through the cultured skin, which resembled in the skin cell homogenates metabolism.

CONCLUSIONCultured at the air-liquid interface, HaCaT cells developed a parakeratotic mutilayer epithelium. Enzyme activity was reserved. This cultured skin could serve as an appropriate model for drug percutaneous metabolism and skin irritation.

Administration, Cutaneous ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacokinetics ; Esters ; administration & dosage ; chemistry ; pharmacokinetics ; HeLa Cells ; cytology ; Humans ; Ketoprofen ; administration & dosage ; chemistry ; pharmacokinetics ; Skin Absorption ; Skin, Artificial ; Tissue Engineering ; methods