OBJECTIVE To investigate the effects and mechanism of luteolin on osteogenic repair of bone defects. METHODS The targets and potential pathways of luteolin in the treatment of bone defects were screened by network pharmacology method， and then the top 2 targets were selected by Hub gene screening for molecular docking verification， with binding energy as the evaluation standard. In vitro experiments were conducted on rat bone mesenchymal stem cells （BMSC） and rat umbilical vein endothelial cells （RUVEC）. Phenotypic validation was performed using alkaline phosphatase staining， alizarin red S staining， and in vitro angiogenesis experiments. Western blot assay was used to detect the protein expressions of phosphatidylinositol 3 kinase （PI3K） and protein kinase 1 （Akt1）， so as to validate the mechanism of luteolin on osteogenic differentiation of BMSC and angiogenesis of RUVEC in vitro. RESULTS The results of network pharmacology showed that the effects of luteolin on vascular formation and bone repair in bone defects were mainly related to Akt1， SRC， estrogen receptor 1， epidermal growth factor receptor， cyclooxygenase 2， matrix metalloproteinase 9 targets， and were closely related to PI3K-Akt signaling pathway. The results of molecular docking showed that luteolin binding to Akt1 and SRC proteins was stable. The results of in vitro experiments showed that luteolin could significantly improve the expressions and activities of alkaline phosphatase in BMSC， increased the number of calcium salt deposits and calcified nodules， and promoted calcification of BMSC. Compared with luteolin 0 μmol/L group， the angiogenesis ability of RUVEC was enhanced significantly in luteolin 1， 10 μmol/L groups， the length of blood vessels and the protein expressions of PI3K and Akt1 were significantly increased （P＜0.05 or P＜0.01）； the higherthe concentration， the better the effect. CONCLUSIONS Luteolin may play a role in promoting angiogenesis and bone repair at the fracture site by activating PI3K/Akt signaling pathway and promoting the protein expressions of PI3K and Akt1.

Chromatin immunoprecipitation sequencing (ChIP-seq) and the Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) have become essential technologies to effectively measure protein–DNA interactions and chromatin accessibility. However, there is a need for a scalable and reproducible pipeline that incorporates proper normalization between samples, correction of copy number variations, and integration of new downstream analysis tools. Here we present Containerized Bioinformatics workflow for Reproducible ChIP/ATAC-seq Analysis (CoBRA), a modularized computational workflow which quantifies ChIP-seq and ATAC-seq peak regions and performs unsupervised and supervised analyses. CoBRA provides a comprehensive state-of-the-art ChIP-seq and ATAC-seq analysis pipeline that can be used by scientists with limited computational experience. This enables researchers to gain rapid insight into protein–DNA interactions and chromatin accessibility through sample clustering, differential peak calling, motif enrichment, comparison of sites to a reference database, and pathway analysis. CoBRA is publicly available online at https://bitbucket. org/cfce/cobra.

The capacity for thermal tolerance is critical for industrial enzyme. In the past decade, great efforts have been made to endow wild-type enzymes with higher catalytic activity or thermostability using gene engineering and protein engineering strategies. In this study, a recently developed SpyTag/SpyCatcher system, mediated by isopeptide bond-ligation, was used to modify a rumen microbiota-derived xylanase XYN11-6 as cyclized and stable enzyme C-XYN11-6. After incubation at 60, 70 or 80 ℃ for 10 min, the residual activities of C-XYN11-6 were 81.53%, 73.98% or 64.41%, which were 1.48, 2.92 or 3.98-fold of linear enzyme L-XYN11-6, respectively. After exposure to 60-90°C for 10 min, the C-XYN11-6 remained as soluble in suspension, while L-XYN11-6 showed severely aggregation. Intrinsic and 8-anilino-1-naphthalenesulfonic acid (ANS)-binding fluorescence analysis revealed that C-XYN11-6 was more capable of maintaining its conformation during heat challenge, compared with L-XYN11-6. Interestingly, molecular cyclization also conferred C-XYN11-6 with improved resilience to 0.1-50 mmol/L Ca²⁺ or 0.1 mmol/L Cu²⁺ treatment. In summary, we generated a thermal- and ion-stable cyclized enzyme using SpyTag/SpyCatcher system, which will be of particular interest in engineering of enzymes for industrial application.
Animals ; Cyclization ; Endo-1,4-beta Xylanases ; chemistry ; metabolism ; Enzyme Stability ; Industrial Microbiology ; methods ; Microbiota ; Protein Engineering ; Rumen ; enzymology ; microbiology ; Temperature

Objective To explore the efficacy of quantitative susceptibility mapping (QSM) in the assessment of osteoporosis and the impact factors on the QSM values.Methods A total of 105 volunteers (35 males and 70 females) were recruited in this study.The height,weight,waistline and hipline were measured,and the body mass index was calculated.All the subjects underwent MRI-based QSM and quantitative computed tomography (QCT).The measurement of QSM and QCT values was performed on L3 vertebrae body.According to QCT value,the subjects were divided into three groups (normal,osteopenia and osteoporosis).According to age,the subjects were divided into group I (21-30 years old),group 2 (31-40 years old),group 3 (41-50 years old),group 4 (51-60 years old),and group 5 (＞60 years old).Differences among all groups were compared using one-way ANOVA or Kruskal-Wallis.Results According to QCT value,54 subjects were normal,22 osteopenic and 29 osteoporotic.The QSM value for the subjects with osteoporosis [148.60(109.42,188.81)ppb] was significantly higher than that of normal (P＜0.001)and the osteopenia (P＜0.001).The QSM value for the subjects with osteopenia was significantly higher than the normal (P＜0.001).The coefficient of QSM and BMD was-0.749 (P＜0.001).Multiple linear regression showed age was the independent influence factor for QSM value (r=0.72,P＜0.001),whereas the gender,BMI,waistline and hipline showed no significant difference (P＞0.05).With the increasing of age,the QSM value showed a gradual increasing trend.And there were significant differences of QSM values among the different age groups (P＜0.001).The QSM value of 138.98 (100.37,183.84)ppb for group 5 (＞60 years old) was significantly higher than that of group 1,group 2,and group 3 (P＜0.001).There is no difference between group 5 and group 4 (P＞0.05).The QSM value of 96.62(28.62,143.99)ppb for group 4 (51-60 years old) was significantly higher than that of group 1 and group 2 (P＜0.001).And there was no difference between group 4 and group 3 (P＞0.05).The QSM value of group 1,group 2,and group 3 showed no significant difference (P＞0.05).Conclusions The QSM of bone is feasible in the assessment of osteoporosis and has the potential to be a biomarker providing new insights into osteoporosis.And age is the critical factor affecting QSM value.

Objective:To explore the correlation between carotid lesion severity detected by ultrasound and pregnancy associated plasma protein A (PAPP‐A) expression in patients with acute coronary syndrome (ACS) .Methods :A to‐tal of 78 ACS patients hospitalized in our hospital from Feb 2012 to May 2015 were regarded as ACS group ,mean‐while ,another 78 healthy subjects were enrolled as healthy control group .Correlation between carotid lesion severi‐ty detected by ultrasound and PAPP‐A expression was analyzed . Results:Compared with healthy control group , there were significant rise in serum PAPP‐A concentration [ (0.97 ± 0.32) mg/L vs .(1.56 ± 0.19) mg/L] ,carotid intima‐media thickness [IMT ,(0.84 ± 0.13) mm vs .(1.28 ± 0.16) mm] and Crouse plaque score [ (2.98 ± 1.92) scores vs .(8.24 ± 1.13) scores] in ACS group ,P<0.01 all .Linear correlation analysis indicated that serum PAPP‐A concentration was significant positively correlated with Crouse plaque score and IMT ( r= 0.342、0.243 , P<0.05 all) .Multi‐factor gradual linear regression analysis indicated that carotid Crouse plaque score and IMT were in‐dependent risk factors for PAPP‐A (partial regression coefficient=1.932 ,17.722 ,P<0.01 both) .Conclusion:Ca‐rotid ultrasound Crouse plaque score ,IMT are significantly positively correlated with PAPP‐A expression ,which can indirectly reflect coronary artery disease severity in ACS patients ,it is worth extending .

Objective: To explore therapeutic effect of recombinant human brain natriuretic peptide (rhBNP) in aged patients with acute decompensated heart failure (ADHF).Methods: A total of 98 aged ADHF patients treated in our hospital were selected and randomly divided into routine treatment group (n=50) and rhBNP group (n=48, received intravenous injection of rhBNP based on routine treatment group).After 72h treatment, cardiac function indexes, total effective rate and incidence of adverse drug reactions (ADR) were measured and compared between two groups.Results: Compared with before treatment, after 72h treatment, there were significant rise in left ventricular ejection fraction (LVEF) and 24h urine volume in both groups(P=0.001 all);compared with routine treatment group after treatment, there were significant rise in LVEF [(45.9±7.8)% vs.(57.4±7.9)%] and 24h urine volume [(1637.5±103.2)ml vs.(1836.4±118.4)ml], P=0.001 all.On 7d after treatment, total effective rate of rhBNP group was significantly higher than that of routine treatment group (93.75% vs.82.00%, P=0.033).There was no significant difference in incidence rate of ADR between two groups, P=0.898.Conclusion: Recombinant human brain natriuretic peptide can significantly improve heart function, and the therapeutic effect is significant in aged patients with acute decompensated heart failure.

Objective: To study influence of glycosylated hemoglobin A1c (HbA1c) on major adverse cardiovascular events (MACE) in patients with diabetes mellitus (DM) complicated coronary heart disease (CHD) after percutaneous coronary intervention (PCI).Methods: A total of 100 DM+CHD patients after PCI were selected from our hospital.According to HbA1c level, they were divided into HbA1c<6.5% group (n=48) and HbA1c≥6.5% group (n=52).Levels of C reactive protein (CRP), tumor necrosis factor α (TNF-α), erythrocyte sedimentation rate (ESR) and interleukin (IL)-6 before PCI, incidence rate of MACE on six and 24 months after PCI were compared between two groups.Results: Compared with HbA1c<6.5% group before PCI, there were significant rise in serum levels of CRP[(18.5±6.2) mg/L vs.(25.8±4.2) mg/L]and TNF-α[(32.4±12.3) ng/L vs.(48.3±11.8) ng/L]in HbA1c≥6.5% group, P<0.01 both.On six months after PCI, incidence rate of myocardial infarction in HbA1c≥6.5% group was significantly higher than that of HbA1c<6.5% group (9.62% vs.0, P=0.028);24 months after PCI, compared with HbA1c<6.5% group, there were significant rise in incidence rates of myocardial infarction (2.08% vs.15.38%) and diseased vessel restenosis (12.50% vs.32.69%) in HbA1c≥6.5% group (P<0.05 all).Conclusion: In DM+CHD patients after PCI, those with lower HbA1c level possess better prognosis.

Objective:To explore therapeutic effect of behavioral nursing on patients with atherosclerotic stenosis after percutaneous coronary intervention (PCI).Methods: A total of 112 patients with atherosclerotic stenosis treated in our hospital were selected.According to ramdom number table and parallel control analysis method, they were randomly and equally divided into routine nursing group and behavioral nursing group (received more active behavioral nursing based on routine nursing group), both groups were treated and observed for six months.The self-care ability scale score before and after nursing, incidence of complications and quality of life (QOL) score after six months were compared between two groups.Results: Compared with before nursing care, after six-month nursing, self-care ability scores significantly rose in both groups, P<0.01 both;compared with routine nursing group after six months, there were significant rise in self-care ability score [(110.34±12.84) scores vs.(128.49±11.39) scores](P=0.001) and each dimension score of QOL(P=0.001 all), and significant reduction in incidence rate of complications (21.4% vs.7.1%,P=0.031) in behavioral nursing group.Conclusion: Behavioral nursing can improve self-care ability, quality of life, promote rehabilitation, effectively reduce occurrence of postoperative complications in patients with atherosclerotic stenosis after percutaneous coronary intervention.

OBJECTIVETo increase local blood supply of bone graft, a novel posterior lumbar spine fusion method with orthotopic paraspinal muscle-pediculated bone flaps was constructed, and the fusion rate and clinical effect.were observed.

METHODSFrom June 2007 to December 2010, 117 patients of lumbar spinal stenosis or lumbar destabilization treated with the novel posterior lumbar fusion method were studied, 49 males and 68 females, aged from 40 to 77 years, average 61.5 years. Clinical effect was evaluated by JOA and VAS score preoperatively and postoperatively, and the fusion result was evaluated by three-dimensional CT reconstruction postoperatively.

RESULTSSeventeen cases lost of follow up, the rest were followed up from 7 to 38 months, average 19 months. There was significant difference between pre- and postoperative JOA and VAS score (P < 0.01), the preoperative JOA score was 10.3 ± 1.9, and 25.4 ± 4.2 at the latest follow-up, the improvement rate was 81.0% ; the preoperative VAS score was 8.5 ± 0.8, and 2.3 ± 0.4 at the latest follow-up. The three-dimensional CT reconstruction showed that 126 of the 133 segments formed solid fusion in 100 patients who completed the follow-up, the fusion rate was 94.7%.

CONCLUSIONThe novel posterior lumbar fusion method make the bone graft position more precise, stable and increases the fusion rate, which can effectively reduce pseudarthrosis and have a promising clinical effect.

Adult ; Aged ; Bone Transplantation ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; Male ; Middle Aged ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery ; Treatment Outcome

Objective To synthesize the hydrophobic supraparamagnetic ironic oxide(SPIO) loaded and hydrophilic SPIO loaded polymeric nano-vesicles and to investigate the feasibility of using hydrophobic SPIO loaded and hydrophilic SPIO loaded polymeric nano-vesicles to display the tumor in MRI in vivo through animal experiments. Methods The polymeric nano-vesicles were prepared from poly (D, L-lactic acid) (PDLLA) and poly (ethylene glycol) (PEG) by a multiple emulsion/solvent evaporation method.The hydrophobic SPIO and hydrophilic SPIO were loaded in the polymeric nano-vesicles respectively.Eighteen nude mice models with human colorectal carcinoma xenograft were established. They were divided equally into three groups (n = 6). The three groups of nude mice models were injected with water-soluble SPIO, hydrophobic SPIO loaded and hydrophilic SPIO loaded vesicle via the mice caudal vein respectively.Dynamic MRI scan were performed in all the mice models. T2WI signal intensity and T2 relaxation time were measured in the tumor, liver and muscle by using T2 mapping software. ANOVA of repeated measurement was used to analyze if there were significant differences of signal intensity changes among the three groups, while Bonferroni method was used for pair-wise comparison. Results On T2 WI, tumors showed decrease in signal intensity after hydrophobic or hydrophilic SPIO loaded polymeric nano-vesicle injection, while no signal intensity decrease was found in the tumor after water-soluble SPIO administration. The maximum percentage of signal intensity decrease in tumor caused by hydrophobic SPIO loaded and hydrophilic SPIO loaded vesicle were 11.00%, 11.40%, respectively. There was statistical significant difference of signal intensity changes among these three groups (F = 10. 96, P ＜ 0. 01). The decrease in signal intensity in the groups with hydrophilic or hydrophobic SPIO loaded polymeric nano-vesicles injection were more pronounced as compared with that of water-soluble SPIO (P ＜ 0. 05), but there was no significant difference in signal intensity decrease between the groups of hydrophilic and hydrophobic SPIO-loaded polymeric vesicles injection (P ＞0. 05). The three agents could lead to signal intensity decrease in the liver. The maximum percentage of signal intensity decrease in liver caused by water-soluble SPIO, hydrophobic SPIO loaded and hydrophilic SPIO loaded vesicle were 32. 85%, 52. 77%, 56. 89%, respectively. There was statistical significant difference between these groups (F = 161.18, P ＜ 0. 01) . The groups of injecting hydrophilic and hydrophobic SPIO loaded polymeric nano-vesicles had the more obvious signal decrease than the one with water-soluble SPIO (P ＜ 0. 01). Hydrophilic SPIO loaded polymeric nano-vesicles exhibited more signal intensity decrease than hydrophobic SPIO loaded polymeric nano-vesicles (P ＜ 0. 01). All three agents could not lead to T2WI signal decrease in the muscle, and there was no significant difference in signal change on T2 WI among three groups (F = 0. 59, P ＞ 0. 05). Conclusion SPIO loaded polymeric nano-vesicles can cause significant T2WI signal loss in human colonic carcinoma on MR imaging in vivo. It can be used as tumor imaging contrast agents.