The high frequency key words in papers on reading therapy published in China were selected, the fre-quencies of co-words presenting in a paper were calculated, and the established matrix of co-words was trans-formed into correlative matrix and diversity matrix, respectively.The diversity matrix was analyzed by cluster analysis and the hot research spots in papers on reading therapy published China were studied.

High frequency key words in papers on MOOC were retrieved from CNKI and Wanfang Data Knowledge Service Platform.A co-words matrix was constructed , which was then transformed into correlation matrix and dis-similarity matrix, respectively.The correlation matrix was analyzed by multidimensional scaling analysis to show the hotspots in domestic MOOC studies on teaching reform, open education, information technology, resource develop-ment, library service innovation and its development trends.

Objective To investigate the the multi-directional differentiation potential between pluripotent of human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLCs). Methods HPDLCs and HGFs were obtained from the primary culture. HPDLCs and HGFs at 3rd-4th passage were cultured in osteogenic, adipogenic or chondrogenic me-dium. Cells without differentiation were taken as control. Alizarin red, Alcian blue and oil red O staining were performed to detect osteogenic differentiation, chondrogenic and adipogenic differentiation in vitro, respectively. Reverse transcription polymerase chain reaction (RT-PCR) was applied to examine the expression of osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and collagen 1 (Col 1), peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and collagen 10 (Col 10). Results HPDLCs and HGFs cultured in osteogenic medium showed massive calicium nodulus at day 28, but HP-DLCs formed more calicium nodulus than those of HGFs. The expressions of OCN, RUNX2 and Col 1 were significantly high-er in HPDLCs than those in HGFs (P<0.05). In chondrogenic medium both cells were found blue deposit at day 14, and the expression of Col 10 was significantly higher in HGFs than that of HPDLCs (P<0.01). Furthermore, in adipogenic medium HGFs showed more lipid-filled droplets stained with oil red O than HPDLCs at day 21. The expression of PPARγ2 was sig-nificantly higher in HGFs than that of HPDLCs (P<0.01). Conclusion HPDLCs has the better potency of osteogenic differ-etiation than HGFs, however, HGFs has the better potency of adipogenic and chondrogenic differentiation.

Cas9 is a RNA-guided double stranded DNA nuclease that participates in the CRISPR/Cas9 system. Wide-type Cas9 directly silences the expression of target gene by gene splicing. The engineered dCas9 protein with the mutation at D10A and H840A lacks the Cas9' s endonuclease function but keeps its DNA binding activity. dCas9 can activate special genes by fusing with transcription activator. Meanwhile,it can inhibit the gene transcription by directly binding to the target gene and stop gene transcrip?tion. Combination of light sensitive structures and CRISPR can produce light-inducible CRISPR/Cas9 system for control of gene expres?sion. This system is able to activate or inhibit gene expression via the use of controlling blue light(470 nm). In this review,we mainly discuss the development of the light inducible CRISPR/Cas9 system as well as its application in the control of gene expression.

Objective To analyze the virulence genes and drug resistance in Vibrio parahaemolyti-cus strains isolated in Nantong City from 2015 to 2016 in order to provide reference for the prevention and treatment of Vibrio parahaemolyticus infection and for rational use of medicines. Methods Virulence genes of tlh,tdh and trh in Vibrio parahaemolyticus strains were detected by fluorescence quantitative PCR. Micro-broth dilution method was used to analyze antimicrobial resistance in these strains to 15 kinds of antibiotics. Results Eighty-two Vibrio parahaemolyticus strains were all positive for tlh gene and negative for trh gene and among them,72 carried tdh gene (87.8%). Antimicrobial resistance rates of these strains to ampicil-lin,cefazolin,tetracycline and chloramphenicol were all 1.2% (1/82). Two strains (2.4%) were resist-ant to trimethoprim/sulfamethoxazole. All strains were sensitive or intermediate to another 10 kinds of antibi-otics. Conclusion From 2015 to 2016,Vibrio parahaemolyticus strains carrying virulence genes of tlh and tdh were prevalent in Nantong and no trh gene-positive strains were reported. Except ampicillin, cefazolin, tetracycline,chloramphenicol and trimethoprim/sulfamethoxazole these five kinds of antibiotics, the remai-ning 10 kinds of antibiotics were effective against Vibrio parahaemolyticus and could be used as the treatment of choice.

Objective To determine whether Tanshinone ⅡA(Tan ⅡA)has an effect on angiotensin Ⅱ(Ang Ⅱ)-induced prolifera-tion and migration of vascular smooth muscle cells(VSMCs),phenotypic switching,or autophagy.Methods In this study,murine aortic smooth muscle cell lines were treated with Ang Ⅱ to establish a model of cell proliferation.Different concentrations of Tan ⅡA were added and effects on cell proliferation and migration were determined by cell counting using a CCK-8 assay and a cell scratch assay,respectively.Alpha-smooth muscle actin(α-SMA),a marker of contractile VSMCs,was detected by Western blotting.Expression levels of osteopontin(OPN)and the autophagy-related proteins(p62,Beclin-1,LC3A/B)were assayed to determine the effect of Tan ⅡA on VSMC phenotypic transformation and autophagy.Cells were treated with the autophagy inhibitor 3-methyladenine(3-MA),combined with Tan ⅡA,and then cell proliferation,migration and expression levels of phenotypic markers and autophagy-related proteins were assessed.Results After Ang Ⅱ treatment,the proliferation and migratory capacity of VSMCs was significantly enhanced,and phenotypic transformation was sig-nificantly inhibited by Ang Ⅱ.Western blotting revealed that Ang Ⅱ reduced the expression of α-SMA,increased the expression of OPN,p62,Beclin-1,and LC3A/B,and that these effects increased with higher Tan ⅡA concentrations.After the addition of 3-MA to the treated cells,the proliferation and migration of VSMCs induced by Ang Ⅱ was inhibited,the expression of α-SMA was increased,and the expres-sion of OPN,p62,Beclin-1,and LC3A/B was decreased,which was consistent with the effect of Tan ⅡA.Conclusion Tan ⅡA may have inhibitory effects on phenotypic transformation,proliferation,migration,and autophagy of Ang Ⅱ-treated VSMC,suggesting that the inhi-bition of the proliferation and migration may be regulated by autophagy.

Objective:To investigate the value of nomograms based on clinical parameters, apparent diffusion coefficient (ADC) and MRI-derived radiomics in predicting survival of patients with locally advanced cervical cancer (LACC) after concurrent chemoradiotherapy (CCRT).Methods:Clinical data of 423 patients with IB-IVA cervical cancer treated with CCRT at Anhui Provincial Hospital Affiliated to Anhui Medical University from March 2014 to March 2020 were retrospectively analyzed and randomly divided into the training and validation groups at a ratio of 2∶1 using the simple randomization method. The values of ADC min, ADC mean, ADC max and 3D texture parameters of diffusion weighted imaging (DWI), T 2WI, T 2WI-fat suppression of pre-treatment primary lesions in all patients were measured. The least absolute shrinkage and selection operator (LASSO) algorithm and logistic regression analysis were used to screen the texture features and calculate radiomics score (Rad-score). Cox regression analysis was employed to construct nomogram models for predicting overall survival (OS) and cancer-specific survival (CS) of patients with LACC after CCRT, which were subject to internal and external validation. Results:Squamous cell carcinoma antigen (SCC-Ag), external beam radiotherapy dose, ADCmin and Rad-score were the independent prognostic factors for OS and CS of LACC patients after CCRT and constituted predictive models for OS and CS. The area under the receiver operating characteristic (ROC) curve (AUC) of two models in predicting 1-year, 3-year, 5-year OS and CS was 0.906, 0.917, 0.916 and 0.911, 0.918, 0.920, with internally validated consistency indexes (C-indexes) of 0.897 and 0.900. Then, models were brought into the validation group for external validation with AUC of 0.986, 0.942, 0.932 and 0.986, 0.933, 0.926 in predicting 1-year, 3-year, 5-year OS and CS.Conclusion:The nomograms based on clinical parameters, ADC values and MRI-derived radiomics are of high clinical value in predicting OS and CS of patients with LACC after CCRT, which can be used as prognostic markers for patients with cervical cancer to certain extent.

Objective:To investigate the value of nomogram based on intravoxel incoherent motion diffusion weighted imaging (IVIM-DWI) and MRI-derived radiomics for predicting recurrence after concurrent chemoradiotherapy (CCRT) in patients with locally advanced cervical cancer (LACC).Methods:Clinical data of 111 patients with ⅠB-ⅣA cervical cancer who underwent CCRT at Anhui Provincial Hospital from December 2014 to December 2019 and were continuously followed up were retrospectively analyzed. Pre-treatment IVIM-DWI parameters (ADC, D, D * and f) and pre- and post-treatment 3D texture parameters (from axial T 2WI) of the primary lesions were measured. Least absolute shrinkage and selection operator (LASSO) algorithm and multivariate logistic regression analysis were used to filter texture features and calculate radiomics score (Rad-score). A Cox regression model was used to analyze independent risk factors for recurrence after CCRT in patients with LACC and construct a nomogram. Results:External beam radiotherapy dose, f value , pre-treatment Rad-score and post-treatment Rad-score ( HR=0.204, 3.253, 2.544, 7.576) were the independent prognostic factors for recurrence after CCRT in cervical cancer patients and jointly formed the nomogram. The area under curve (AUC) of the nomogram for predicting 1-, 3- and 5-year disease-free survival (DFS) was 0.895, 0.888 and 0.916, with internal validation C-indexes of 0.859, 0.903 and 0.867, respectively. The decision curves analysis showed that the nomogram has a higher net clinical benefit compared to other models, and the clinical impact curves further visualized its predictive accuracy. Conclusions:The nomogam based on IVIM-DWI and radiomics has high clinical value in predicting recurrence after CCRT in patients with LACC, providing reference for prognostic assessment and individualized treatment of cervical cancer patients.

Objective To evaluate the therapeutic effect of evodiamine(Evo)on atopic dermatitis(AD)in rat models by regulating the cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)/cAMP response ele-ment binding protein(CREB)signaling pathway.Methods A rat model of AD was established by administration of multiple doses of 2,4-dinitrochlorobenzene(DNCB).The animals were randomly divided into AD group,Evo-low-dose(Evo-L,5 mg/kg)group,Evo-high-dose(Evo-H,10 mg/kg)group,Evo-H+H-89(5 mg/kg)group and dexamethasone(0.1 mg/kg)group.Normal rats were used as the control group and then the degree of skin damage of rats in each group was scored.Abdominal blood was taken to detect the levels of interleukin-4(IL-4),cAMP,and tumor necrosis factor-α(TNF-α)in serum;Skin lesion tissue was collected to detect pathological change,counting of mast cells,PKA/CREB related protein expression and expression of IL-4 and TNF-α mRNA in the tissue.Results Compared with control group,the level of cAMP in serum,the expression of p-PKA/PKA,and p-CREB/CREB in skin lesions of AD group were reduced,the severity score of skin lesions,level of IL-4 and TNF-α,epidermal thickness,number of mast cells and mRNA expression of IL-4 and TNF-α in skin lesion tissues were all significantly increased(P＜0.05).Compared with AD group,the level of cAMP in serum,the expression of p-PKA/PKA,and p-CREB/CREB in skin lesions in Evo-L group,Evo-H group,and dexamethasone group were increased,the severity score of skin lesions,level of IL-4 and TNF-α,epidermal thickness,number of mast cells,and mRNA expression of IL-4 and TNF-α in skin lesion tissues all reduced(P＜0.05).Compared with the Evo-H group,the level of cAMP in serum,the expression of p-PKA/PKA,and p-CREB/CREB in skin lesions in Evo-H+H-89 group was reduced and the severity score of skin lesion,level of IL-4 and TNF-α,epidermal thickness,num-ber of mast cells,and mRNA expression of IL-4 and TNF-α in skin lesion tissues significantly increased(P＜0.05).Conclusions Evo inhibits inflammatory response and pathological damage through regulation of cAMP/PKA/CREB signaling pathway in AD rat models.

Animals ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Mice ; Plasmids ; RAW 264.7 Cells ; RNA, Guide