To screen the pathogenic mutation location in a genetic family with the neurofibromatosis (NF1) by the next generation sequencing and analyze the clinical phenotype,Illumina Miseq sequencing was applied to capture and analyze the target regions of NF1 family's probands,and furtherly find out the suspicious mutations,as well as to verify the family members by Sanger sequencing.Two rare variants were identified in proband,including the heterozygous missense mutation c.C3649T (p.P1217S) in KIF1B gene and the missense mutation c.T6311C (p.L2104P) on exon 41 of NF1 gene (NM_000267.3).The amino acid at position 2104 was found to be changed from leucine to proline in NF1.The protein prediction SIFT and Polyphen-2 values were 0,0.997,which predicted a conformational change in the encoded protein and eventually affected its function.The mutation c.T6311C in NF1 gene was detected in all patients in this family,which showed genetic co-segregation.The clinical phenotype was neurofibroma in the spinal canal.There were no café au lait spots,iris Lisch nodules,scoliosis,tinnitus,heating loss,or elevated intracranial pressure.The missense mutation c.T6311C (p.L2104P) in NF1 gene might be the genetic cause of this hereditary disease of neurofibromatosis.

BACKGROUND:In recent years,some scholars in the field of tendon bone injury have attached stromal cell-derived factor 1 to tissue engineering scaffolds to promote tendon bone healing,and achieved good results.However,whether stromal cell-derived factor 1 promotes tendon bone healing mechanisms and participates in the repair of natural healing has not yet been defined. OBJECTIVE:To study the expression of stroma-cell derived factor 1 during tendon bone healing after rupture of the whole supraspinatus muscle of the rabbit rotator cuff and its migration effect and optimal in vitro migration promoting concentration on stem cells during tendon bone injury. METHODS:Totally 18 adult New Zealand rabbits were randomly selected to establish rotator cuff injury models,and an additional 3 rabbits were selected as blank controls.At 3,5,7,14,21,and 28 days after modeling,three rabbits were executed separately and the rabbits in the blank group were sacrificed.The tissues of tendon bone junction were taken and stored in a-80℃refrigerator.The expression of stromal cell-derived factor 1 was detected by ELISA at each time point after injury.Mesenchymal stem cells were isolated from the bone marrow of young rabbit femur,cultured,and identified.Transwell assay was performed to verify the migration-promoting effect of stromal cell-derived factor 1 on stem cells and the optimal migration-promoting concentration in vitro.The stem cells cultured to P3 were co-cultured with BrdU and injected into the rabbit ear marginal vein,and immunohistochemical staining was used to verify whether the stem cells migrated to the injury site. RESULTS AND CONCLUSION:(1)Stromal cell-derived factor 1 gene expression was bimodal during rotator cuff tendon bone healing.Stromal cell-derived factor 1 gene expression increased significantly at 3 days post-injury(P<0.01)and then decreased,reaching a minimum at 5 days post-injury.It increased again and reached a peak 14 days after injury(P<0.01)and then decreased.(2)Cell immunohistochemical staining displayed that stem cells labeled with BrdU did migrate to the injury site.(3)The results of the transwell experiment exhibited that 60-80 ng/mL stromal cell-derived factor 1 had the best effect on promoting migration of stem cells,while a concentration of 200 ng/mL inhibited migration.(4)Stromal cell-derived factor 1 is involved in the healing of rotator cuff tendon bone during the inflammatory response phase and the proliferation phase.The mechanism of action may be to promote the migration of stem cells to the injury and their differentiation into various types of cells to promote repair.In addition,the pro-migration effect of stromal cell-derived factor 1 exists at a range of concentrations,beyond which it may act as an inhibitor.

OBJECTIVE：To explore the potential active ingredients and mechanism of Alpinia officinarum in the treatment of gastric ulcer. METHODS ：By network pharmacology method ，the active ingredients and action targets of A. officinarum were screened through TCMSP and TCMID database retrieval [oral bioavailability （OB）≥30％ and drug like （DL）≥0.18] and literature mining. Targets of gastric ulcer were obtained in the TTD ，CTD，OMIM，PubMed，DrugBank and DisGeNet databases. Venny 2.1 software was used to screen common targets for the active ingredients of A. officinarum and gastric ulcer. Then ，the protein-protein interaction（PPI）of the common targets was obtained by STRING database ，and the PPI network was constructed and analysed by using Cytoscape 3.7.1 software. GO function and KEGG pathway enrichment analysis of the common targets were performed by using ClusterProfiler R package. Finally ，Cytoscape 3.7.1 software was used to construct and analyze the network diagram of “active ingredients-targets-pathways ”. RESULTS ：Totally 19 active ingredients of A. officinarum ，209 active ingredients targets and 195 gastric ulcer related targets ，involving 35 common targets ，were screened out. The average node degree of PPI network of common targets was 18，and the average intermediate number was 16.9. There were 11 key targets ，i.e. PTGS2，VEGFA，IL6， IL1B，CCL2，MYC，MMP9，EGFR，HIF1A，ESR1，BCL2L1. The common targets were mainly concentrated in the cell constituents such as the platelet α granule lumen and mitochondria outer membrane ，involved in the biological processes as oxidative stress ，inflammatory response regulation ，and molecular functions as protein phosphatase binding ，growth factor receptor binding. They were also enriched in the signal pathways such as PI3K/Akt，HIF-1. The network of“active ingredients- targets-pathways”showed the active ingredients such as quer- cetin，apigenin，kaempferol and galangin in A. officinarum played an anti-gastric ulcer effect by acting on PTGS2，NOS2， BCL2， IL6， VEGFA， IL1B， MMP9， BCL2L1 and other targets to jointly regulate PI 3K-Akt，HIF-1，TNF，IL-17， NF-κB and other cell proliferation，angiogenesis，and infla- 163.com mmation related pathways. CONCLUSIONS ：A. officinarum shows anti-gastric ulcer effect with the characteristics of multi-ingredient ，multi-target and multi-path.