From 38 pieces of deep\|sea samples,a strain,DY\|A,which produces protease was isolated.It can grow at a wide range of pH and salt concentration and best at 10℃.The strain produced maximum protease activity after growth at 10℃,initial pH 10 0 and inoculation volume 0 5%for 48～72h on a shaker by speed of 200r/min.The optimum pH and temperature for the protease activity production are pH 10.0 and 40℃.The enzyme is alkaline protease.Some other properties of the strain and the protease were discussed in detail.

[Objective]To investigate the effect of a single subcutaneous dose of an antisense oligonucleotide(ASO) on particle-induced osteolysis.[Method]The murine calvaria osteolysis model was utilized in ICR mice.Bone resorption was measured with the toluidine blue staining.Osteoclasts were detected by tartrate resistant acid phosphatase(TRAP) staining assay and were quantified by a TRAP quantification kit.[Result]Bone resorption was 0.347 ? 0.09 mm2 in animals with particle implantation,and decreased to 0.123 ? 0.05 mm2 and 0.052 ? 0.02 mm2 after ASO treatment in low and high doses,respectively.The bone resorption was reestablished in animals given an additional TNF-?.The number of osteoclasts in animal calvaria treated with ASO was reduced obviously compared with those untreated animals and the quantification results indicated that about 90% osteoclastgenesis was suprressed by the ASO.Additionally,the osteoclastgenesis was reestablished by the addition of TNF-?.[Conclusion]An antisense oligonucleotide targeting an inflammatory factor,TNF-?,has been to suppress the osteolysis induced by particle for the first time.This new finding holds a great promise.It is a therapeutic strategy for the component loosening.

Objective To report the preliminary results of Less Invasive Stabilization System (LISS) applied in treatment of proximal tibial fractures. Methods 22 patients with acute fractures of proximal tibia were treated with LISS plates from November, 2003 to June, 2004. There were 6 females and 16 males, with a mean age of 41.8 years (ranging from 22 to 69 years). According to AO classification, 7 cases were type 41- A, and 15 cases were type 41- C (9 combined with other fractures). 5 of these were open fractures (3 cases of Gustilo- Anderson type Ⅲ A and 2 cases of type Ⅲ B). Results All fractures were eventually healed. There were 2 cases of superficial infection. No deep infection, failure of fixation or malalignment was found in this group. However, 1 case showed displacement of articular surface 6 months postoperatively. The range of motion was from 2? to 98.6? . The mean time of radiographic healing was 14.5 weeks (ranging from 11 to 17 weeks) and the mean time of full weight bearing was 15.2 weeks (ranging from 12 to 18 weeks). By Johner - Wruhs' evaluation, 13 cases were rated as excellent, 7 as good, 2 as fair 1 year postoperatively. Conclusions LISS is a safe and effective fixation device for proximal tibial fractures, especially for those caused by high- energy injuries. The benefits of this system include less intervention of blood supply, less soft tissue complications and a high union ratio of fractures.

Objective To explore the roles of IκBα and TGF-β1 on airway inflammation in rats with chronic bronchitis induced by smoking and study the effects and mechanism of anti-inflammation of pretreatment with ambroxol. Methods Sixty male wistar rats were randomly divided into four groups: Normal dosage group, model group, high dosage group and low dosage group. The rats with chronic bronchitis were established by smoking. For high and low dosage group, rats were pretreated respectively with ambroxol group, rats were pretreated with normal saline through peritoneal injection as much as the low dosage group.After 76 days, the histopathologic changes stained in hemotoxylin and eosin (H. E.) of bronchopulmonary tissues were observed under opticalmicroscope, white cell counts and differential analysis were performed in BALF, the expression of IκBα and TGF-β1 were detected by immunohistochemistry. Results The pathological changes of model group were in consistent with that of chronic bronchitis, but the degrees were significantly reduced in high and low dosage groups. Compared with those in normal group, the white cell count and the neutrophilic granulocyte count of BALF in model group were significantly increased and the macrophagocyte count decreased, and the expression of IκBαwas significantly decreased(t =3.24,3.31,3.29,3.48, P ＜0. 05) and the TGF-β1 significantly increased (P ＜0. 05). Compared with those in the model group, the white cell count and the neutrophilic granulocyte count of BALF in high and low dosage group were significantly decreased and the macrophagocyte count increased, the expression of IκBαwas significantly increased (t =2. 86,2. 97,2. 92,3.52,2.42,2. 88,2. 58,3.48, P ＜0. 05) and the TGF-β1 significantly decreased (P ＜0. 05) . Compared with those in low dosage group, the expression of TGF-β1 was decreased and the expression of IκBαincreased in high dosage group (t =2. 82,3.64, P ＜0. 05). Conclusions Down-regulating the expression of IκBα and up-regulating of TGF-β1 may be involved in the process of airway inflammation in rats with chronic bronchitis induced by smoking. Ambroxol might have better effects on ameliorating airway inflammation by up-regulating the expression of IκBα and down-regulating of TGF-β1.

Objective To investigate the correlation between the polymorphism of XRCC2 gene,a DNA double-strand break repair(DDSBR)gene,and the susceptibility to lung cancer in population of North China.Methods By PCR-based restriction fragment length polymorphism(RLFP)technique(PCR-RLFP),a case-control study was performed among 300 patients with lung carcinoma and 300 healthy controls to detect XRCC2(C41657T)polymorphism.Results There was no significant difference in XRCC2(C41657T)allele frequencies between lung cancer patients and healthy controls(P=0.16).The C/C,C/T and T/T genotype frequencies were 72.7%,24.0% and 3.3% in lung cancer patients,and 76.3%,22.0% and 1.7% in healthy controls,respectively.There was no significant difference in C/C and C/T genotype frequencies between cancer patients and healthy controls(?2=0.48,P=0.48).However,the C/T genotype frequency in non-smoke group was significantly higher in lung cancer patients than in healthy controls(?2=6.67,P=0.01).The risk for lung cancer in non-smokers was 2.11 times higher in C/T genotype carriers than that in C/C genotype carriers.There was no significant difference in C/C and C/T genotype frequency between lung cancer patients and healthy controls in the smoke group(P=0.16).Conclusion Overall,there was no significant correlation between genetic polymorphism of XRCC2(C41657T)and the susceptibility to lung cancer.But the C/T genotype might increase the risk of suffering from lung cancer in no-smoking populations of Northern China.

The effects of imperatorin (Imp) and iso-imperatorin(Isi) on tumor necrosis factor (TNF) release from mouse peritoneal macrophages were investigated. It was found that Imp and lsi significantly inhibited TNF release from mouse peritoneal macrophages. At the concentration of 10-6～10-4 mol?L-1, the inhibitory effects were presented by Imp and Isi in a dose dependent manner. At 10-4 mol/L TNF release was entirely inhibited by each drug.

Objective To study the molecular biological mechanism of the protective effect of angelica on myocardium during ischemia reperfusion Methods The myocardial ischemia/reperfusion was induced with the 40 min cross clamp/120 min declamping of anterior decending coronary artery Forty SD rats were randomly divided into 3 groups: group A without myocardial ischemic reperfusion , group B with intravenous adminisrtration of normal saline 0 8ml/100g before ischemia reperfusion , and group C with intravenous adminisrtration of 25% angelica 0 8ml/100g before ischemia reperfusion The content of protein kinase C (PKC) in cardiac myocyte was measured with immunohistochemical method, and the PKC activity with the isotope lable method Results Compared with those in group B, the myocardial infarct size reduced significantly in group C (P0 05) but increased obviously in group C (P0 05) The PKC activity was significantly higher in group C than that in group A(P

Objective: To establish an enzyme linked immunosorbent assay(ELISA)for erythropoietin(EPO) in serum, and observe its clinical application value thereof. Methods: Prepare the EPO polyclonal antibody, wash the plate with isopropyl alcohol, and then choose the suitable concentration of the antibody, enzyme labeled antibody, and antigen. After the reaction, check the sensitivity, recovery, specificity and stability of the method. The serum samples of anaemia and breast carcinoma individuals were detected. The results of radioimmunodetection were compared with that of normal control group. Results: The immo-assay plate showed strong adherence to proteins. The optimal concentrations of the antibody, enzyme labelled antibody and antigen were 1∶1 000, 1∶6 000 and 1∶800 separately. The sensitivity was 0.46 U/L. The cross-reaction with growth hormone and ferritin was low. The mean recoveries of samples with high and low concentrations were 96.3%, 97.3% respectively. The coefficients of variation of intra-assay and inter-assay were just 8.31% and 7.82%, and the stability was good. The EPO levels were higher in anaemia and breast carcinoma groups than that of normal group. There was no significant difference between the results of the radioimmunodetection and ELISA. Conclusion: The double-antibody sandwich ELISA method was established for EPO in serum, which had certain clinical application value.

Phosphorylation of beta-catenin tyrosine residue 654 plays an important role in the epithelial to myofibroblast transition (EMT). Introducing mimic peptide of tyrosine residue 654 domain of beta-catenin into cells may influence phosphorylation of beta-catenin tyrosine residue 654. To deliver this mimic peptide into renal epithelial cells, we used penetratin as a vector, which is a novel cell permeable peptide, to deliver hydrophilic molecules into cells. A tyrosine 654 residue domain mimic peptide of beta-catenin (PM) with fused penetratin was constructed, purified and then detected for the penetration of the mimic peptide into human renal tubular epithelial cells (HK-2). The results showed that purified fusion mimic peptide could efficiently and rapidly translocate into human renal tubular epithelial cells. It is concluded that a cell-permeable peptides mimic of tyrosine residue 654 domain of beta-catenin was successfully obtained, which may provide a useful reagent for interfering the human renal tubular epithelial-mesenchymal transition.
Carrier Proteins/*metabolism ; Epithelial Cells/cytology ; Epithelial Cells/*metabolism ; Fibroblasts/cytology ; Fibroblasts/metabolism ; Kidney Tubules/*cytology ; Peptides/metabolism ; Permeability ; Phosphorylation ; Tyrosine/*metabolism ; beta Catenin/*metabolism

Objective To explore the relationship between prenatal diagnosis indications and fetal chromosomal aberrations , and the security of amniocentesis. Methods The amniotic fluid cells were sampled by amniocentesis and cultured in 572 high-risk pregnant women from January 2012 to August 2015. The chromosomal karyotypes were examined by G-banding. Results The success rate of the first amniotic fluid cells culture reached 99.83%. In all the 572 valid samples , there were 20 cases of chromosomal aberrations and the abnormal rate was 3.50%, including 17 of numeric aberrations and 3 of structural aberrations. There were 7 cases of chromosomal aberrations in all the 299 elderly parturient in high-risk indications and the abnormal rate was 2.34%, and there were 13 cases of chromosomal aberrations in all the 273 non-elderly parturient and the abnormal rate was 4.76%. Conclusions (1)It is necessary to further diagnose in pregnant women with high-risk factors , including high age , abnormal screening and ultrasonic findings , and history of abnormal gestation and birth. (2)The chromosomal karyotype examination of amniotic fluid cells in high-risk pregnant women is one of the effective prenatal diagnosis methods in high security and accuracy , with which it can reduce the incidence of birth defects and the burden of family and society , and improve the quality of the population.