Objective To clone acidic mammalian chitinase (AMCase) gene and analyze the sequences of the nucleotides and encoded amino acids. Methods Total RNA was isolated from the stomach of BALB/c mouse and the mRNA was reversely transcribed into cDNA. AMCase gene was amplified by polymerase chain reaction and inserted into recombinant clone vector of pMD18-T. Sequence alignment was performed with DNAMAN 4.0 software. Isoelectric point, amino acid composition, signal peptide and transmembrane helices were analyzed with SignalP 3.0 Server and TMHMM Server v.2.0. Results The cloned gene encodes a protein of 473 amino acids with molecular weight of 51.93 kDa and isoelectric point 4.73. Extensive amino acid sequence homology (up to 99.57%) was found when compared with AMCase. Both protein encoded by this cloned gene and AMCase belonged to family 18 chitinase, which was composed of signal peptide (amino acid position 1-21), the catalytic domain (amino acid position 22-391), hinge region (amino acid position 392-425) and chitin binding domain (amino acid position 426-473). There were two mutations of amino acid in the catalytic domain and hinge region with this AMCase mutant. The amino acid of Ala(position 292)was mutated into amino acid of Pro, and the amino acid of Thr(position 404)was mutated into amino acid of Ser. Conclusion This sequence is a mutant of AMCase gene. Identification of the biological characteristics will play important role in the further study of its biological function by molecular biology technique.

We developed a three-dimensional finite element model of the shoulder glenohumeral joint after shoulder arthroplasty including humerus shaft, scapular, scapular cartilage and eight muscles, while each of the muscles was simulated with 50 spring elements. To reduce the element number and improve the analytical precision, we used mixed tetrahedral and hexahedral elements in the model. We then used the model to calculate the biomechanics of the shoulder glenohumeral joint after hemiarthroplasty during humeral external rotation. Results showed that the maximum joint reaction force was 374.72 N and the maximum contact stress was 6. 573 MPa together with the contact areas at 40 degrees external rotation. These might be one of the reasons for prosthetic disarticulation, and would provide theoretical bases to prosthetic design.
Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Humeral Head ; anatomy & histology ; Models, Anatomic ; Prosthesis Design ; Range of Motion, Articular ; Rotation ; Shoulder Joint ; anatomy & histology

Identifying the similarity and differences of malpractice,medical negligence and medical error has realistic meaning to deal with medical disputes efficiently.They are all subjective negligence and belong to the category of medical behavior.This paper mainly discussed the criteria and civil liability of their differences.

Objective To study the potential roles of acoustic immittance,distortion product otoacoustic e-mission (DPOAE),and the brainstem response audiometry (ABR)in the diagnosis of children with acute non-sup-purative otitis media (AN-SOM).Methods A retrospective analysis was administered to 182 cases with AN-SOM at the otolaryngology department between March 2014 and March 2015.We compared the results of the acous-tic immitance of 218 symptomatic ears and 146 asymptomatic ears,and also the proportion of DPOAE abnormity and ABR abnormity.We analyzed the specialty of symptomatic ears and asymptomatic ears in 3 terms of audiology test results.Results The proportions of abnormal tympanograms,DPOAE,ABR for symptomatic ear were 64.7%, 72.0%,and 57.8%,which were significantly higher than those of in 41.8%,39.7%,and 35.6% in asymptomatic ear.The difference were statistically significant(P<0.05).A further analysis of audiologic results showed that at least one abnormal result was as high as 196 ears (89.9%)in the asymptomatic ear,90 ears (61.6%)showed no symptoms.The differences were statistically significant(P<0.05).The results of audiology tests showed that tym-panograms in symptomatic ears showed diversity,that were 60.6% for type B or C,35.3% for type A,4.1% for the As.Conclusion The early detections by using acoustic immitance,DPOAE and ABR are very important to the diagnosis of children with acute non-suppurative otitis media.It can improve the sensitivity,and reduce the misdi-agnosis.

Objective To clone and express the mouse acidic mammalian chitinase ( AMCase ) gene and protein, and to analyze the hydrolysis of the cell walls of Trichophyton rubrum and Candida a/ft/cans by this enzyme. Methods Total RNA was isolated from the stomach of BALB/c mouse, and AMCase gene was amplified by RT-PCR. The recombinant fusion expression vector of pET28a ( + ) -AMCase was constructed. AMCase protein was expressed in prokaryotic system and purified. After incubation of AMCase with the cell wall extracts, the level of N- acetylglucosamine was measured. Results AMCase gene was cloned and expressed successfully. Purified AMCase protein can hydrolyse the chitin in the cell walls of T. rubrum and C. albicans. Conclusion The AMCase expressed in prokaryotic system can hydrolyse chitin in the cell walls of T. rubrum and C. albicans, which implies its antifungal potential.

Objectives To evaluate the count of endothelial progenitor cells (EPCs) in peripheral blood (PB) and bone marrow (BM) of acute leukemia (AL) patients and explore its clinical significance.Methods EPCs were detected by flow cytometry procedures in 43 AL patients and in 10 benign hematologic patients as control group.Results The absolute counts of EPCs in AL patients before the treatment [(119.46± 72.23)/μl in BM and (13.69±8.26)/pl in PB] were significantly higher than those in control group [(23.21 ± 12.59)/pl in BM and (1.86±1.18)/μl in PB] (P ＜ 0.01).The absolute counts of EPCs were significandy higher in BM than those in BP in AL patients before the treatment (P ＜ 0.001).After the treatment, the absolute counts of EPCs in no remission (NR) group [(110.02±67.28)/μl in BM and (10.04±9.51)/μ1 in PB] were significantly higher than those in control group (P ＜ 0.05), while the counts of EPCs in complete remission (CR) group were no significant difference compared with those in control group (P ＞ 0.05).After the treatment ,the absolute counts of EPCs both in BM and in BP of CR group [(26.32±17.44)/μl and (2.54±2.12)/μl, respectively] were significantly lower than those before treatment [(113.18±69.22)/μl and (14.45±10.76)/μl, respectively] (P ＜ 0.05), however those of NR group were no significant difference than before (P ＞ 0.05).The absolute counts of EPCs whether in PB or in BM were no significant difference between acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL) (P ＞ 0.05).The absolute counts of EPCs in PB of AL had a positive correlation with β2-MG and LDH (P ＜ 0.05).Conclusions EPC levels are significantly increased in BM and BP of AL patients and may correlate with disease status, response to treatment and prognosis.

Objective To investigate the serum levels of advanced oxidation protein products (AOPP) in patients with chronic obstructive pulmonary disease (COPD) as well as explore the relevance of its clinical significance AOPP and COPD.Methods Fifty-four patients with mild/moderate COPD (COPD group) were enrolled in this study,who were treated in the No.100th Hospital of the Chinese People's Liberation Army from Apr.2011 to Nov.2013.Thirty healthy volunteers (control group) at same period were selected as control group and the general condition of two groups were matched.AOPP,superoxide dismutase(SOD) and malondialdehyde (MDA) levels were measured.Results The serum AOPP,MDA and SOD in control group were (45.78 ± 12.54) μmol/L,(2.96 ± 0.55) μmol/L and (78.40 ± 8.37) kU/L respectively.And its were (68.93 ± 10.62) μmol/L,(6.07 ± 2.44) μmol/L and (53.66 ± 5.99) kU/L respectively in COPD group.The differences were statistically significant (t =-8.57,-9.14,14.38 ; All P values were less than 0.01).The serum AOPP,MDA and SOD in mild COPD group were (65.56 ±9.65) μmol/L,(4.21 ± 1.83) μmol/L and (62.97 ± 6.28) kU/L respectively,and (71.79 ± 11.37) μmol/L,(7.43 ± 3.12) μmol/L and (41.25 ± 5.89) kU/L respectively in moderate CODP group.The differences were statistically significant (t =-2.17,-4.80,13.00; P ＜ 0.05 or P ＜ 0.01).Conclusion The increased levels of serum AOPP is important pathological changes in patients with COPD,which may be involved in the occurrence and development of COPD,and is the reaction of oxidative stress injury in patients with COPD early sensitive indicator.

Objective AIM:To study whether the RWD containing sumoylation enhancer (RSUME) enhanced small ubiquitin related modifiers (SUMO) to competitively inhibit Ubiquitin B (UBB)-mediated degradation of hypoxia hypoxia-inducible factor 1α.(HIF-1α) and the invasive pituitary adenomas.Methods The expression of protein and mRNA levels of RSUME,SUMO-1,UBB and HIF-1α were detected by using immunohistochemistry,western blot and qPCR in 38 cases of non-invasive pituitary adenoma,38 cases of invasive pituitary adenomas and 10 cases of normal pituitary capsule.The expression of SUMO-1 was analyzed in different types of pituitary adenomas.Results The protein,and mRNA levels of RSUME,SUMO-HIF-1αwere significantly higher in the invasive pituitary adenomas than in the non-invasive pituitary adenomas and normal pituitary capsule (P＜0.01).The protein and mRNA levels of RSUME,SUMO-HIF-1αwas higher in non-invasive pituitary adenomas than in normal pituitary capsule(P＜0.01).However,the protein levels of UBB-HIF-1α were significantly lower in the invasive pituitary adenomas than in the non-invasive pituitary adenomas and normal pituitary capsule (P＜0.01).The protein of UBB-HIF-1α were lower in non-invasive pituitary adenomas than in normal pituitary capsule (P＜0.01).The expression levels of SUMO were not significantly different among different types of pituitary adenomas (P＞0.05).Conclusion RSUM may increase pituitary adenomas angiogenesis and promote tumor invasion through enhancement of SUMO of HIF-1α which competitively inhibits ubiquitination of HIF-1 α.

BACKGROUND:We have found that oriented fibers can guide the alignment of smooth muscle cells in our previous experiments. Thus, we designed the experiment to prepare wel aligned polymeric fibers using electrospinning technology, aiming at guiding the growth of esophageal smooth muscle cells to maintain cellmorphology and biological function. OBJECTIVE:Using electrospinning technology, to fabricate isotropic and directed nano-fibrous scaffolds made of polycaprolacton, gelatin and silk fibroin. METHODS:Polycaprolacton/silk fibroin fibers at a ratio of 4:1 were prepared with proper parameters, including solution concentration, voltage and injection speed, under the self-made spinning system. The polycaprolacton/gelatin sheets with mass ratio of 2:1, 1:1 and 1:2, respectively, were also fabricated under suitable process parameters. Using the rol er col ector instead of the metal plate, polycaprolacton/gelatin nano-fibrous scaffold with good alignment of fibers was manufactured. RESULTS AND CONCLUSION:The isotropic polycaprolacton/silk fibroin scaffold with fiber diameter of (535.9±126.7) nm was prepared under conditions of solution concentration (0.08 g/mL), injection speed (1.6 mL/h) and voltage (22.5 kV), and these fibers were uniform with no beads. The isotropic polycaprolacton/gelatin scaffold with fiber diameter of (257.9±117.8) nm was prepared under conditions of solution concentration (0.10 g/mL), injection speed (0.8 mL/h) and voltage (22.5 kV). Using the rol er col ector instead of the previous metal plate, polycaprolacton/gelatin (w:w, 1:2) nano-fibrous scaffold with good alignment of fibers was manufactured. The process parameters were 3 000 r/min of rol ing speed, 0.8 mL/h of injection speed and 15 kV of voltage.

In contemporary studies of pulse and of the relationship between sphygmo-diagram and syndrome types or diseases, pulse instrument is usually applied to trace the sphygmo-diagram. A comparatively systemic theory about pulse diagnosis has been formed, and it promotes the pulse researching process. But the mechanism of pulse is complicated and the expressive information of pulse is diversity. So it is difficult to record the complicated information of pulse by applying the instrument. In addition, the simplicity in methods of tracing and analyzing sphygmo-diagram and the lack of criterion for syndrome differentiation make it difficult to study the relationship between the sphygmo-diagram and the syndrome types. It's important to lay stress on clinical applying, to promote communication among researchers, to unify the standards of pulse instrument and syndrome differentiation, and to reinforce the research on the relationship between sphygmo-diagram and the syndrome types. The government's support is also needed to promote multi-science cooperation at the same time.