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.

OBJECTIVETo evaluate the short-term efficacy and long-term preventive effect on recurrence in prevention and treatment of children allergic rhinitis treated with the multiple therapy at Dazhui (GV 14) as the main acupoint and acupoint plaster therapy.

METHODSOne hundred and twenty cases of children allergic rhinitis were randomized into a triple-strong stimulation therapy group and an acupoint plaster therapy group, 60 cases in each one. In the triple-strong stimulation therapy group, Dazhui (GV 14) was the main acupoint, combined with one of Feishu (BL 13), Zhiyang (GV 9) and Shenshu (BL 23) accordingly. The heavy needling technique was adopted with the three-edged needle, followed by heavy cupping and heavy moxibustion. The treatment was given once every 3 days, continuously for 10 times. In the acupoint plaster therapy group, the acupoints selected were same as the triple-strong stimulation therapy group. The self-prepared Chinese herbal plaster was used, 4 h to 8 h each one time, once every 3 days, continuously for 10 times. The changes in the symptom and physical sign scores were observed before treatment, after treatment and in 6 months follow-up visit after treatment in the two groups. The clinical efficacy was compared between the two groups.

RESULTSAfter treatment and in 6 months follow-up visit after treatment, the symptom scores, physical sign scores and symptom scores of each item were different signi ficantly as compared with those before treatment in the triple-strong stimulation therapy group (P<0.01, P<0.05), and the results in the triple-strong stimulation therapy group were better than those in the acupoint plaster therapy group (all P<0.01). In 6 months follow-up visit, the total effective rate was 94.6% (53/56) in the triple-strong stimulation therapy group and was 25.9% (15/58) in the acupoint plaster therapy group, indicating the significant difference (P<0.001).

CONCLUSIONThe triple-strong stimulation therapy at Dazhui (GV 14) achieves the superior effect on the prevention and treatment of children allergic rhinitis as compared with the acupoint plas ter therapy and has good long-term effect of the prevention from recurrence.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Child ; Combined Modality Therapy ; Female ; Humans ; Male ; Moxibustion ; Rhinitis, Allergic ; prevention & control ; therapy

Objective To investigate the association between carotid atherosclerosis(AS) and serum magnesium(Mg) in hemodialyzed (HD) patients.Methods Clinical index was measured,and intimamedia thickness (IMT) of extracranial common carotid artery and presence of atherosclerotic plaques were determined by high-resolution B-mode ultrasonography.The data were analyzed between plaque positive group and plaque negative group.Results The age,serum phosphate (P),total cholesterol (TCH),low density lipoprotein (LDL),serum C-reactive protein (CRP),serum albumin(Alb),and serum Mg all had significant difference between two groups(t =4.153,2.908,2.301,6.322,5.791,2.341,7.778,P ＜0.01 or P ＜ 0.05).The risk factors of HD patients with AS were serum Mg and CRP(P ＜ 0.01).Conclusions The occurrence of AS was related to low Mg and high CRP in HD patients.

BACKGROUND:Heart valve surgery combined with coronary artery bypass grafting has been gradual y used for treatment of coronary heart disease combined with cardiac valve disease.
OBJECTIVE:To retrospectively summarize the experience of combined coronary artery bypass grafting and valvular procedure.
METHODS:Total y 51 patients who underwent combined heart valve surgery and coronary artery bypass grafting were retrospectively analyzed, including 10 cases with aortic valve replacement, 14 cases with mitral valve replacement, eight cases with mitral valve replacement combined with tricuspid annuloplasty, four cases with aortic valve and mitral valve replacement, three cases with aortic valve and mitral valve replacement combined with tricuspid annuloplasty, seven cases with mitral valvuloplasty, and five cases with mitral valvuloplasty combined with tricuspid annuloplasty.
RESULTS AND CONCLUSION:Biovalve replacement was performed in nine patients, mechanical valves replacement in 31 cases, mitral valvuloplasty in 11 cases and tricuspid annuloplasty in 16 cases. There were total y 109 bypass graft vessels, and the average number of coronary artery bypass grafts was (1.92±0.73) branches. Four cases died within 30 days postoperatively, and 47 patients were successful y discharged from the hospital. Forty-five of 47 discharging patients were fol owed for 3-48 months. One case died of cerebral infarction within 6 months postoperatively, and another case died of cardiac dysfunction over 1 year after operation. The heart function of 45 survival patients was significantly improved. The comprehensive analysis showed that improving the heart function preoperatively, strengthening myocardial protection, shortening operation and myocardial ischemia time, and complete revascularization are the key factors for successful operation.

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.

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

AIM To investigate the effect of aspirin on the proliferation and NOS expression in astrocytoma cell line, and probe into its mechanism. METHOD The effect of aspirin on the growth of astrocytoma cells was evaluated by MTT assay; NOS protein levels were determined by immunocytochemistry, NO and CEA concentration in the medium were determined by Griess assay and lepton catch immunising method respectively. RESULTS Aspirin inhibited the growth of astrocytoma cells, induced the expression of iNOS, increased the concentration of NO in the medium. The effects of these were centratoin dependent. Moreover, aspirin reduced the concentration of CEA in the medium. CONCLUSION Aspirin inhibits the growth of astrocytoma cell line. Up regulated iNOS expression resulting a increase of NO concentration are ascribed to mechanism of antiproliferation activity of aspirin. CEA is a good indicator in monitoring curative effect of astrocytoma.

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 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.

BACKGROUND:The methods to treat intertrochanteric fracture are controversial. A large number of clinical studies concern the therapeutic effects of several popular methods to repair intertrochanteric fracture, but these results lack of independence, and may have bias that cannot be measured in the variable and observational studies. Thus, relevant studies have been limited. OBJECTIVE:To compare the repair effects of proximal femoral nail anti-rotation, dynamic hip screw and total hip arthroplasty on intertrochanteric fracture in aged patients by meta-analysis. METHODS: Four electronic databases were searched, including Cochrane library, PubMed, Web of Science and Chinese BioMedical Literature Database, to colect al randomized controled trials concerning the treatment of intertrochanteric fractures in the elderly with proximal femoral nail anti-rotation, dynamic hip screw and total hip arthroplasty. Using meta-analysis of evidence-based medicine, X-ray exposure time, postoperative complication rate, postoperative bed time, postoperative wound infection rate, Harris hip scores, mean operation time, intraoperative blood loss, length of hospital stays and length of the incision were compared and evaluated. Standard and methodology quality of the trials were criticaly assessed and relative data were extracted. This study used the Review Manager 5.0 software provided by Cochrane colaboration network. RESULTS AND CONCLUSION:Twelve randomized controled trials with 1 454 patients were included. Significant differences in mean operation time, intraoperative blood loss, X-ray exposure time, postoperative complication rate, and postoperative bed time were detected between the proximal femoral nail anti-rotation and dynamic hip screw groups (P < 0.05). Significant differences in length of hospital stays, postoperative complication rate, and postoperative bed time were observed between the proximal femoral nail anti-rotation and total hip arthroplasty groups (P < 0.05). Significant differences in postoperative complication rate, postoperative bed time, and Harris hip scores were detectable between the dynamic hip screw and total hip arthroplasty groups (P < 0.05). These data confirm that proximal femoral nail anti-rotation was apparently better than dynamic hip screw and total hip arthroplasty in operation time, intraoperative blood loss, length of hospital stays, postoperative complication rate, and postoperative bed time. Dynamic hip screw was better than proximal femoral nail anti-rotation in X-ray exposure time. Total hip arthroplasty was better than dynamic hip screw and proximal femoral nail anti-rotation in length of hospital stays, postoperative complication rate and postoperative bed time.