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.

BACKGROUND:Although bone tissue engineering scaffolds made of traditional methods have made certain achievements, the three-dimensional structure, mechanical strength and personalized property of the scaffolds are unsatisfied. 3D printing technology is expected to change these shortcomings. OBJECTIVE:To review the 3D printing of bone tissue engineering scaffolds and to prospect the optimization of the scaffolds. METHODS:A computer-based search of PubMed and Google academic database was performed for articles addressing the 3D printing of bone tissue engineering scaffolds published from 2008 to 2015. Articles concerning the structure design and materials of bone tissue engineering scaffolds and different 3D printing technologies for scaffold preparation were included, and repetitive and old articles were excluded. Final y, 37 articles were summarized. RESULTS AND CONCLUSION:Currently, 3D printing technologies used for preparation of bone tissue engineering scaffolds include melt laminated molding, stereolithography, selective laser sintering and 3DP technology. 3D printing technologies have unique advantages in mechanics, structure and personalized aspects, but there are stil many problems to be solved, such as raw materials, insufficiency of different 3D technologies, and improvement of 3D printer. Under the multi-disciplinary co-operation, 3D printing technology is expected to prepare suitable bone tissue engineering scaffolds and bring benefit to the mankind.

Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.

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 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.

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 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.

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 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.

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.