ABSTRACT
Objective To summarize clinical characteristics and investigate possible pathogenic gene of Klippel-Feil syndrome(KFS)by the self-designed multigene panel sequencing,so as to decipher the molecular basis for early diagnosis and targeted therapy.Methods From January 2015 to December 2018,we consecutively recruited 25 patients who were diagnosed with KFS in Peking Union Medical College Hospital.The demographic information,clinical manifestations,physical examination and radiological assessments were analyzed.Multigene panel sequencing was performed after DNA extraction from peripheral blood.The possible pathogenic mutations of KFS were explored on the basis of bioinformatics analysis.Results The KFS cohort consisted of 25 patients,including 15 males and 10 females,with a mean age of(12.9±7.3)years.Limited cervical range of motion was the most common clinical feature(12 cases,48%).Based on the Samartzis classification,the proportion of patients suffered from short neck(P=0.031)and limited cervical range of motion(P=0.026)in type Ⅲ KFS was significantly higher than that in type Ⅱ and type Ⅰ KFS.Panel sequencing detected a total of 11 pathogenic missense mutations in eight patients,including COL6A1,COL6A2,CDAN1,GLI3,FLNB,CHRNG,MYH3,POR,and TNXB.There was no pathogenic mutation found in five reported pathogenic genes(GDF6,MEOX1,GDF3,MYO18B and RIPPLY2)associated with KFS.Conclusions Our study has shown that patients with multiple contiguous cervical fusions are more likely to manifest short neck,limited cervical range of motion,and clinical triad.Therefore,these patients need additional attention and follow-up.Our analysis highlights novel KFS-related genetic variants,such as COL6A and CDAN1,extending the spectrum of known mutations contributing to this syndrome and providing a basis for elucidating the pathogenesis of KFS.
Subject(s)
Child , Female , Humans , Male , Cervical Vertebrae , Cohort Studies , Glycoproteins , Klippel-Feil Syndrome/genetics , Mutation , Nuclear Proteins , Radiography , Transcription Factors/geneticsABSTRACT
ObjectiveTo investigate the effects of extracts ofLiaoxuan Kaxifu Powders (LE) on proliferation and transcription of IL-8 and ICAM-1 in HaCaT induced by TNF-α; To discuss its mechanism of action.Methods Cultured HaCaT was assigned into normal group, TNF-α group and low-, medium- and high-dose of LE group. Every group was induced by 40 ng/mL TNF-α except for normal group, and then LE groups were treated by different concentrations (8, 40, 200μg/mL) of LE for 48 h. The proliferation of HaCaT was evaluated by MTT and the apoptosis was detected by inverted fluorescence microscope. Levels of IL-8 and ICAM-1 in HaCaT were assessed by ELISA, and their mRNA expressions was detected by semi-quantitative RT-PCR.Results Compared with normal group, the absorbency of HaCaT and the contents and mRNA expressions of IL-8 and ICAM-1 increased in TNF-α group (P<0.05,P<0.01); compared with TNF-α group, LE of all dose groups could significantly inhibit the absorbency, decrease the contents and mRNA expressions of IL-8 and ICAM-1 (P<0.05,P<0.01).Conclusion LE is able to inhibit the proliferation of HaCaT induced by TNF-α, and the mechanism is probably related to promoting apoptosis and down-regulating the gene expressions of IL-8 and ICAM-1, and then maintaining the normal level of HaCaT.
ABSTRACT
Objective To study the producing method for Euphorbia pekinensis promoted by endophytic fungi.Methods The tissue culture plantlets were innoculated with endophytic fungi,then it was transplanted to the pods and cultured for one year.The biomass and content of terpenoids(isoeuphpekinensin and euphol)were determined.Results The growth of the E.pekinensis was promoted by endophytic fungi E4(Fusarium sp.Ⅰ)and E5(Fusarium sp.Ⅱ).The fresh weight of the root increased 49.45% and 59.74% for E4 and E5 treated,respectively.The plants dry weight coefficient increased 2.99% and 6.48% for E4 and E5 treated,respectively.The contents of isoeuphpekinensin and euphol in the plants were increased for tissue cultured in both bottles and pods.For the plants cultured in pods for one year,the isoeuphpekinensin and euphol increased 92.79% and 40%,respectively treated by E4.The isoeuphpekinensin and euphol increassed 105.32% and 241.38%,respectively treated by E5.Conclusion The endophytic fungi E4 and E5 could promote the growth of E.pekinensis and accumulated terpenoids.