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.

Objective To explore the whole quality control method for lactones and flavonoids in the raw material of Herb Sarcandra.Methods TLC identification of ethyl acetate extracts and spectrophotometry determination of total flavonoids had been conducted.Results Chemical compositions extracted by ethyl acetate and total flavonoids content varied greatly in the raw materials of Herb Sarcandra from different producing areas.Conclusion TLC identification combined with total flavonoids determination can be used for the quality control of the medicinal material of Herb Sarcandra.

Purpose:To study the RNA interference-mediated inhibition of survivin gene on the proliferation of human breast cancer SKBr-3 cells. Methods:SKBr-3 cells were transfected with a pSUPER-S1 vector plasmid that expressed survivin-targeted small interfering RNA, and the mRNA and protein levels of survivin gene were measured with RT-PCR and indirect immunofluorescence, respectively. The proliferation of transfected SKBr-3 cells was investigated through colony forming assay and MTT assay. The cell cycle phases were determined by flow cytometry(FCM). Results:The mRNA and protein levels of survivin declined markedly in pSUPER-S1-transfected SKBr-3 cells. And the colony forming rate of those cells(38?16.70)% was significantly lower than that of the control cells(90.3?4.04)%.The growth of the pSUPER-S1-transfected cells was decelerated and the cell cycle was mainly blocked at G_(1) phase(74.03?8.91)%. Conclusions:survivin gene silencing by RNA interference contributed to a distinctive inhibition of the proliferation of human breast cancer SKBr-3 cells in vitro.

Scientific innovation teams are key weight indicators and promoters in science and technology of a hospital.The First Affiliated Hospital of Anhui Medical University(the Hospital)draws funding support from the Natural Science Foundation,to empower its innovative teams with programs and funding.These efforts have effectively propelled its innovations spearheaded by the dermatology team to give rise to outstanding outcomes and power teams.The application for talents support from NSF,the project approval for the hospital funding is affected to some extent.With the help from NSF and the innovative teams,scientific teams of the hospital have harvested a general upgrading and accelerated development of disciplines,elevating the scientific level of hospital in general.

Objective To study the inhibition of AP-1 transcription factor activity by Hint1 gene over expression in HepG2 cell lines. Methods The Hintl gene was amplified, and then was inserted into the pcDNA3/HA eukaryotic expression plasmid. The constructed pHA-Hint1 plasmid was confirmed by DNA sequencing. The pHA-Hint1 was transfected into the HepG2 human hepatoma cells. Semi-quantitative RT-PCR and Western-blot were used to detecte the expression of HA-Hint1. The HepG2 cells were co-transfected with pHA-Hint1 and pAP-1/Luc luciferase reporter. At 36 h after transfection, luciferase assay system was used to detect the AP-1 transcription factor activity. Results The constructed pHA-Hint1 was confirmed by DNA sequencing, pHA-Hint1 gene transduction through lipofectine induced over-expression in HA-Hint1 mRNA (t =3.89, P<0.05) and HA-Hintl protein (t=3. 12, P<0.05). Co-transfection of Hint1 gene inhibits AP-1 luciferase activity. Cotransfection with increased concentration of a pHA-Hint1 plasmid (0 μg/ml, 0. 5 μg/ml, 1.0 μg/ml, 1.5 μg/ml, 2. 0 μg/ml) produced a concentration-dependent inhibition of AP-1 transcription factor activity. At the concentration of 1.5 μg/ml, and 2.0 μg/ml, the activity inhibition reaches significant difference ( F = 72. 009, P < 0. 05 ). Conclusion Over-expression of Hintl can, at least in part, inhibit the AP-1 transcription factor activity in HepG2 cells.

Objective To investigate the changes of serum adiponectin level and insulin resistance in patients with obstructive sleep apnea hypopnea syndrome (OSAHS).Methods 80 adult habitual snorers were divided into mild,moderate,severe OSAHS groups and control group,based on their apnea hypopnea index (AHI).Correlative clinical data were measured and analyzed.Results Serum adiponectin level [( 5.44 ± 2.57 ) mg/L] was significantly lower and HOMA-insulin resistance index ( 3.42 ± 0.70) was significantly higher in severe group than other groups ( P ＜ 0.05 ).Serum adiponectin level was negatively correlated with BMI,waist hip ratio,neck circumferences and et al( r =-0.233 ～ -0.499,P ＜0.05),but it was positively correlated with lowest oxygen saturation ( LSaO2 ),Ⅲ + Ⅳ％ and REM％ ( r =0.270～ 0.429,P ＜ 0.05).Conclusions Both hypoadiponectinemia and insulin resistance were observed in severe OSAHS patients.The related risk factors of hypoadiponectinemia in OSAHS might include obesity,disordered sleep architecture and other adipokines.

Objective To observe the clinical efficacy of electroacupuncture in treating deglutition disorders after radiotherapy of nasopharyngeal carcinoma. Method Forty nasopharyngeal carcinoma patients with deglutition disorders after radiotherapy were randomized into a treatment group and a control group, 20 in each group. The control group was intervened by nourishing nerve, improving microcirculation, promoting nerve growth factor, and physical rehabilitation. Based on the treatment given to the control group, the treatment group was additionally intervened by electroacupuncture. Water drinking test was adopted to evaluate the deglutition function before and after intervention, and the clinical efficacy was compared. Result After intervention, there was no significant difference in comparing the deglutition function between the two groups (P>0.05). The total effective rate was 80.0%in the treatment group versus 50.0% in the control group, and the difference was statistically significant (P<0.05). Conclusion Electroacupuncture is an effective method in treating deglutition disorder after radiotherapy of nasopharyngeal carcinoma.

Objective summarize the delicacy management practices through the NSFC application procedures in a university affiliated hospital,to provide further reference for improvement of the NSFC project approvai ratio.Methods To summarize the management experiences by analyzing the approval number,project category and funds of NCFS from 2006 to 2015.Results The delicacy man agement practices include cultivate scientific research atmosphere by strengthen motivation,initiate application as early as possible,enlarging application number by extensive mobilization,improve applica tion quality by massive training,multi-round updates,tutorial,prevent avoidable errors by cross-over review.Conclusions Delicacy management by scientific research management department during the application process is critical to improve the project approval ratio of NCFS in a hospital.

Whole mount in situ hybridization with BHC80 RNA probe showed that BHC80 was expressed in zebrafish central nervous system. Morpholino-modified antisense oligonucleotide was injected into zebrafish zygotes to knock down BHC80 expression. BHC80 knockdown resulted in striking decrease of erythrocytes and accumulation of erythrocytes at PBI. Further investigation of embryonic erythrocytes marker βe3 globin and hematopoiesis transcription factors gata1, c-myb and lmo2 by in situ hybridization showed that the erythroid progenitors marked with gata1 in BHC80 knockdown embryos were high proliferation and their differentiation were delayed, which led to decrease of erythrocytes and accumulation of erythrocytes at PBI. Both in situ hybridization and microangiography indicated that vasculature pattern of BHC80 knockdown embryos were almost normal.

Objective: To evaluate the efficacy of Tuina (Chinese therapeutic massage) manipulation plus horse-riding squat exercise in treating knee osteoarthritis (KOA) and optimize the combining protocol. Methods: Based on a 2×2 factorial design, 120 eligible KOA patients were randomized into a manipulation group (group A1B2), a manipulation plus horse-riding squat group (group A1B1), a sitting knee-adjustment group (group A2B2 group), and a sitting knee-adjustment plus horse-riding squat group (group A2B1), with 30 cases in each group. The intervention was conducted three times a week, lasting for four weeks. The Western Ontario and McMaster Universities osteoarthritis index (WOMAC) was taken as the major measure for efficacy evaluation (including three component scores, pain, stiffness, and daily function, and total score). Results: The three component scores (pain, stiffness, and daily function) and the total score of WOMAC showed significant differences after the intervention in the four groups (P<0.05). There were significant inter-group differences in the WOMAC stiffness score amongst the four groups after the intervention (P<0.05). In group A1B1, the step length, stride, walking speed, and knee joint flexion angle changed significantly after treatment (P<0.05). After the intervention, the step length changed significantly in group A1B2 (P<0.05), and the walking speed changed significantly in group A2B1 (P<0.05). There were no significant differences in the step length, stride, walking speed, or knee joint flexion angle among the four groups (P>0.05). The extensor peak torque at 180 °/s changed significantly in group A1B2 after treatment (P<0.05). Neither the intra-group nor the inter-group comparisons of the four groups revealed significant differences in the other isokinetic muscle strength parameters (P>0.05). The main effect of manipulation showed significant in affecting the WOMAC pain and total scores (P<0.05). The main effect of horse-riding squat exercise showed significant in affecting the WOMAC pain and stiffness scores (P<0.05). Conclusion: The four treatment protocols all can improve the symptoms of KOA, for instance, relieving pain and stiffness, and enhancing daily function. Group A2B1 produces the most eminent effect in relieving joint stiffness. The main effects of both manipulation and horse-riding squat exercise are significant in reducing pain. Besides, the main effect of horse-riding squat exercise is significant in relieving joint stiffness.