Two sets of primers were designed according to the sequences of xynA and xynB from Aspergillus sulphureus, and the DNA fragments composed of 574 bp and 594 bp were amplified by polymerase chain reaction (PCR), respectively. The two PCR products respectively digested with EcoRⅠ/BamHⅠ and BglⅡ/HindⅢ were ligated into multiple cloning sites of pET-28a(+). The resulting plasmid is pET-xynAB, in which xylanase A and B are ligated by a 7-amino acid peptide (GlyGlyGlySerGlyGlyGly). E. coli BL21 transformed with pET-xynAB was induced by IPTG, and a special protein band about 50 ku was detected by SDS-PAGE. The protein purified with Ni-NTA column was denatured by 8 mol/L urea and dialyzed for refolding. The recombinant xylanase showed optimal activity at 50℃ and pH 4.4. The enzyme retained above 75% of its activity at the range of pH 2.4～5.4. The xylanase displayed about 50% retained acitivity after incubating at 80℃ for 30 min. Various metal ions have different effects on activity of the recombinant xylanase.

Object To assess the genetic diversity among ten natural populations in Ophiopogon japonicus (L f ) Ker Gawl and O. bodinieri Levl (Liliaceae) in Sichuan Province Methods Forty random decamer primers were screened for Random Amplified Polymophic DNA (RAPD) fragments Results Based on cluster analysis of 515 DNA bands amplified by 11 primers, a DNA molecular dendrogram was established, and the relationship of the populations and seeds between O japonicus and O bodinieri was set up Conclusion 1 Distinct genetic differences and extensive genetic diversity are presented among the samples The genetic differences are related to morphological differences, but not to geographic regions; 2 Quite genetic differences are presented between O japonicus and O bodinieri, it suggests that they have farther relationships

Objective Previous study found that ifbroblast growth factor-2(FGF2) expression was at a low level in bone marrow of children with aplastic anemia. In this study, we established a bone marrow failure animal model to investigate whether FGF2 is involved in bone marrow failure. Methods BALB/c mice were irradiated by 5.0 Gy ray, and then infused with 1×106 lymphocytes from allogeneic mice lymph node. Peripheral blood cells and bone marrow mononuclear cells counting and bone marrow pathology were done. FGF2 protein in bone marrow mononuclear cells was measured by ELISA. Results Compared with the control group, the counting of hemoglobin, white blood cell, platelet and bone marrow mononuclear cell in aplastic anemia mouse model were signiifcantly deceased (P<0.05). Moreover, FGF2 expression in aplastic anemia mouse model were signiifcantly reduced (P<0.05). Conclusions 5.0 Gy ray irradiation and then 1×106 lymphocyte infusion in mice can induce bone marrow failure similar to the features of aplastic anemia. The low expression of FGF2 in bone marrow of aplastic anemia patients may play an important role in the pathogenesis of aplastic anemia.

Objective To assess the effects of 670nm LED (light-emitting diode) to protect the photoreceptor from the light-induced damage in a rat model. Methods 32 SD rats were randomly assigned to one of eight groups: untreated control group, the LED-treated control group, three groups of light-induced damage,and three groups of light-induced damage treated with LED. Light-induced damage result from exposing to constant light for 3 hours of different illuminations of 900,1800 and 2700 lx, respectively. The LED treatment (50 mW) was delivered for 30 minutes at 3 hours before the light damage and 0,24 and 48 hours after the light damage. Retinal function and morphology were measured by electroretinogram (ERG) and histopathology assay. Results The illumination of 900 lx for 3 hours did not damage the rat retina. The illumination of 1800 lx for 3 hours resulted in thinner ONL and no OS and IS. The ratio of damaged area/total retinal area was 0.48±0.12, the damaged thickness of ONL/normal ONL (L5) was 0.39±0.07,and the amplitude of ERG b wave was (431±120) μV. With the LED treatment the ratio of damaged area decreased (M6=0.17±0.12, P5/6=0.002), and the ratio of the damaged thickness of ONL also decreased (L6=0.22±0.09, P5/6<0.01), and the amplitude of ERG b wave increased to (1011±83) μV(P5/6 <0.001). The illumination of 2700 lx for 3 hours caused severed damage to the rat retina and the LED could not protect them significantly. Conclusions 670 nm LED treatment has an evident protective effect on retinal cells against light-induced damage, which may be a simple and effective therapy to prevent or to delay age-related maeular degeneration.

Objective To explore whether Forkhead O transcription factor-3a (FoxO3a) activity affects the capability of sphere-formation of ovarian cancer SKOV3 cell line.Methods Sphere-forming cells (SFCs) were obtained and amplified through suspended culture with conditioned medium of the stem cells in SKOV3 cell line.After SKOV3 cells were transfected with FoxO3a specific siRNA,the protein expressions of FoxO3a and Bmi1 and the ratio of sphere-formation were compared with Western blot and sphere-forming assay,respectively.Results Compared to parental cells,SFCs from SKOV3 cell line had higher ratio of sphere-formation and over-expressed Bmi1 and pFoxO3a.Transfection of FoxO3a specific siRNA down-regulated the protein expression of FoxO3a and upregulated expression of Bmi1 in SKOV3 cells,and enhanced the capability of sphere-formation.Conclusions Silence of FoxO3a leads to enhanced capability of sphere-formation in SKOV3 cell line.

Objective To analyze and assess the major functions of medical APP by investigating its application in self management of chronic diseases. Methods The 3 major functions of APP were analyzed, including data record and condition assessment, medical consultation, and life therapy. Results The medical APP possesses its basic functions for chronic diseases management. However, it lacks of practicality and convenience and its homogenization is severe. Conclusion The APP +hardware +cloud medical management model should be promoted by making use of the Internet + medical technology, the supervision of mobile medical APP should be strengthened, the mobile information management mechanisms of chronic diseases should be perfected, and the self management efficiency of patients with chronic diseases should be improved.

Objective To develop an HPLC method for the determination of ginsenoside Rb1 in Jupizhuru decoction,and de-termine the amount of ginsenoside Rb1 in 10 batches of Jupizhuru decoction composed of different batch herbs. Methods The HPLC analysis was carried out on a Phenomenex Luna C18 column with column temperature at 30℃,flow rate 1.0 ml/min,detection wave-length 203 nm and mobile phase acetonitrile and 0.1%phosphoric acid by gradient elution. Results The method was found to be lin-ear within the ranges of 5-500μg/ml(r=0.9999),and the average recovery of ginsenoside Rb1 was 95.5%(RSD 1.77%,n=6). Con-clusion The determination method is suitable for the quality control of ginsenoside Rb1 in Jupizhuru decoction.

Mitochondria, the power house of cells, are important organelles in eukaryotic cells. Having their own unique and complete DNA (mtDNA) and genetic system, mitochondria play an essential role in cellular energy metabolism, intracel?lular signaling and apoptotic pathways, as well as many other biological functions, which are closely related with cellular met?abolic network. A disruption of mitochondrial genes can therefore result in mitochondrial dysfunction and human diseases, thus they have been widely used in molecular biology, development biology, genetics, forensic identification and clinical diag?nosis. Consequently, sequencing mitochondrial genome has shown great significance in mitochondrial structure and function research. In this review, research progress in mitochondrial genome sequencing method is summarized, mainly focusing on Sanger sequencing, long-PCR and next-generation sequencing. Also rolling circle amplification and indirect sequencing of mtDNA are reviewed. The ambiguities caused by numts in indirect sequencing are mentioned and resolved.

Objective To evaluate the effects of sevoflurane postconditioning on mitophagy during myocardial ischemia-reperfusion (I/R) in rats.Methods Forty-two pathogen-free adult male SpragueDawley rats, weighing 250-300 g, were randomly divided into 3 groups (n=14 each) using a random number table: sham operation group (group S) , I/R group and sevoflurane postconditioning group (group SP).Myocardial I/R was induced by 30 min ligation of the left anterior descending branch of the coronary artery followed by 2 h of reperfusion.In group SP, 2.4％ sevoflurane was inhaled for 15 min starting from the onset of reperfusion, while 33％ oxygen was inhaled in group I/R.The rats were sacrificed at the end of reperfusion, and the hearts were removed for measurement of myocardial infarct size (by 1％ 2, 3, 5 triphenyltetrazolium chloride) , expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1, p62 and Parkin (by Western blot) ,and mitochondrial menbrane potential (by using JC-1 probe) , and for examination of the uhrastructure of cardiomyocytes (with transmission electron microscope).Results Compared with group S, the myocardial infarct size was significantly increased, mitochondrial membrane potential was decreased, the expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1 and Parkin was up-regulated, and the expression of p62 was down-regulated in group I/R.Compared with group I/R, the myocardial infarct size was significantly decreased, the mitochondrial membrane potential was increased, and the expression of LC3 Ⅱ/LC3 Ⅰ , Beclin-1, p62 and Parkin was down-regulated in group SP.Conclusion Sevoflurane postconditioning can mitigate I/R injury in rats, and inhibition of excessive activation of mitophagy may be involved in the nechanism.

Objective To investigate the relationship between the echinoderm microtubule associated protein like 4-anaplastic lymphoma kinase (EML4-ALK) and epithelial growth factor receptor (EGFR) mutation status and overall survival (OS) in Uygur patients with stageⅣnon-small cell lung cancer (NSCLC) who did not accept tyrosine kinase inhibitor treat-ment. Methods Totally 97 tissue samples were collected from Uygur patients with stageⅣNSCLC who did not accept tyro-sine kinase inhibitor treatment. EML4-ALK fusion gene and EGFR mutation status were detected by using FISH and ARMS methods. The survival rates were analysed. Results In 97 tissue samples, EML4-ALK fusion genes were found in 6 (6.2%) samples, EGFR mutations were found in 26 (26.8%) samples. The survival analysis showed that there was no significant dif-ference in OS between EML4-ALK fusion gene group and no EML4-ALK fusion gene group (P=0.941). There was no signifi-cant difference in OS between EGFR mutation group and wild-type EGFR group (P=0.607). The values of median OS were 17.7 months, 17.3 months and 16.2 months for EGFR mutant group, EML4-ALK positive group and EML4-ALK negative+EGFR wild-type group, and thre was no significant difference between them (P=0.915). Conclusion Excluding the thera-peutic influence in TKIs, EML4-ALK fusion gene and EGFR mutation status of tumor tissue can not be used as an indepen-dent factor in assessing the prognosis in Uygur patients with stageⅣNSCLC.