Human FceR I a subunit extracellular domain was cloned and expressed with 2 different expressing systems and Dot blot was used to detect its biological activity to bind with IgE,providing reference for binding mechanism of human FceR I a subunit extracellular domain with IgE. It was shown that FceR I a subunit extracellular domain from pBAD/g I A expressing system could bind with IgE, but the one from PQE30 expressing system could not bind with IgE. It is suggested that FceR I a subunit extracellular domain alone is sufficient to bind with IgE without P and Y subunit. The proper space configuration and disulfide bond of FceR I a subunit is necessary for binding with IgE, but its glycosylation is unnecessary.

Objective To determine the surface expression of Fas Ag on RBL 2H3 and its function. Methods RT PCR and Western blot were used to detect the transfection and expression of Fas in RBL 2H3. Surface expression of Fas Ag was studied by immunochemistry. Apoptosis changes following treatment with anti Fas antibody were analyzed using flow cytometic analysis with annexinⅤ. Results It was successful in amplifying gene of rat Fas Ag, and a band of 32kD was detected by Western blot. The Fas Ag expression on the surface of RBL 2H3 by immunochemistry. RBL 2H3 exhibited apoptosis in response to anti Fas treatment. Conclusion Induction of rat mast cell apoptosis by activation of the Fas pathway provide the mechanism by which the number of mast cells may be regulated in the potential therapeutic strategy for anaphylactic diseases

ObjectiveTo study the histologic changes of contracture Achilles tendon under Ilizarov distraction technique.MethodsEighteen dogs were installed ilizarov external fixation frames on the right hind leg in a neutral position.Pull traction started 1 week postoperatively with speed of 1 mm/day for 3 weeks.After 15 degrees of foot drop,the fixation was still maintained for 3 weeks.Nine animals were randomly selected and put to death(Contracture group),other animals corrected for 3 weeks with the same speed and methods,then put to death(Force group),health specimens as control group.(1)Observe tendon cells arranged distribution and collagen fiber form with HE dyeing,and analysis of tendon retinal thickness with image analysis software.(2)Observe collagen Ⅰ and Ⅲ with picrosirius red dying,and analyzed both content changes with image analysis software.(3)Determination of protein contents of polysaccharide with phloroglucinol spectrophotometry.Results (1)In the contracture group,the collagenous fibre gap was not obvious,the tendon cell assumes spindle-shaped,saccate,the collagen fibrous bundle distortion,the arrangement was scattered in disorder,difficult to see the intercellular substance.Tendon retinal thickness showed obvious differences with the health side (Control group).In the force group,the tendon cells were round or oval,located in collagen bundles in a string of alignment,inflammatory cells were seen.Tendon retinal thickness showed no differences with the health side.(2)With polarized light microscopy,the collagen Ⅰ and collagen Ⅲ composition proportion in the contracture group showed obvious differences with the control group,force group had not obvious differences with health side.(3)The content of polysaccharide in the contracture group were obviously reduced,while force group returned to normal level.ConclusionThe tissue form of contracture Achilles tendon can recovered to normal with the Ilizarov technology.

Objective To identify the specific microRNA (miRNA) that can be taken as a molecular marker for human bone marrow-derived mesenchymal stem cells (MSCs). Methods MSCs were isolated and cultured from bone marrow through density centrifugation and then were induced to differentiate into osteoblasts and chondrocytes. Samples of MSCs, osteoblasts and chondrocytes were detected by miRNA microarrays single channel fluorescence chip to determine the expression levels of miRNAs. Significance Analysis of Microarrays ( SAM, version 2.1 ) software was used to analyze the raw data to determine the miRNAs overexpressed in MSCs, which was validated in the same sample using real time reserve transcriptase polymerase chain reaction (RT-PCR). Results MSCs were successfully isolated from bone marrow and induced to differentiate into osteoblasts and chondrocytes in vitro. Microarrays showed that eight miRNAs (has-miR-424, has-miR-34a, has-miR-593, has-miR-10a, has-miR-148a,has-miR-602, mmu-miR-709 and mmu-miR-665) were overexpressed in MSCs but underexpressed in osteoblasts. Three miRNAs including has-miR-424, PREDICTED_MIR189 and mmu-miR-665, were overexpressed in MSCs but underexpressed in chondrocytes. The has-miR-424 expression in MSCs was 6.6times higher than in osteoblasts and 4.4 times higher than in chondrocytes. The results of real time RTPCR showed that the miR-424 was overexpressed in MSCs, 3.6 times higher than that in osteoblasts and 3.1 times higher than that in chondrocytes, which was coincident with the results of microarray. Conclusions The screened MSCs express more miRNAs in comparison with osteoblasts and chondrocytes,play important roles in maintaining self renewal and undifferentiation of MSCs and is a promising specific molecule marker for MSCs.

Objective:To explore the effect and mechanism of miRNA sponge on the epithelial-mesenchymal transition (EMT) of gastric carcinoma cell lines SGC7901. Methods:Synthetic ZEB2 3'UTR plasmid and siRNA targeting ZEB2 were transfected into the SGC7901 cell line by Lipofectamine 2000. Real-time quantitative polymerase chain reaction was performed to evaluate the expres-sion levels of miR-200a/b/c. Finally, the migratory, invasive, and proliferative activities of the gastric carcinoma cells in vitro were ana-lyzed by the scratch test, the Transwell cell invasion, and the cell cloning assay. The expression of the target protein was detected by Western blot. Results:Compared with the control group, the expressions of miR-200a/b/c significantly decreased, and their migration, invasion, and proliferation capabilities were considerably higher after they were transfected with ZEB2 3'UTR. Although the expres-sions of miR-200a/b/c significantly increased, the migratory, invasive, and proliferative activities of SGC7901 cells also degraded after they were transfected with siRNA targeting ZEB2. The expression of ZEB2 increased, and that of E-cadherin decreased at the protein level after they were transfected with ZEB2 3'UTR. The protein expression of Vimentin in SGC7901 cells significantly increased. The indicators show the opposite trend when cells were transfected with siZEB2, and the differences between the control and mutation groups were insignificant. Conclusion:ZEB2 3'UTR can regulate EMT course by regulating the miR-200a/b/c expression in gastric car-cinoma, consequently regulating the invasion and migration of carcinoma cells.

Objective To explore the effects of zinc finger E-box binding protein (ZEB)2 3′UTR gene transfection on proliferation, invasion and migration in human gastric epithelial cell line GES-1. Methods The synthetic ZEB2 3′UTR and miR-200b micmics were transfected into GES-1 cell line by lipofectamine 2000. We set up control grop, the mutation group and ZEB2 3′UTR group. Real-time quantitative PCR was performed to evaluate the expression levels of miR-200a/b/c and ZEB1/ZEB2 mRNAs after transfection.And then we set up control group, ZEB2 3′UTR group, ZEB2 3′UTR+negative control group and ZEB2 3′UTR+miR-200b micmics group. The protein expression levels of ZEB1, ZEB2, matrix metallopro-teinases (MMP) 2/9 and proliferating cell nuclear antigen (PCNA) were detected by Western blot assay. The invasion and mi-gration capability were analyzed by transwell assay and wound healing test. MTT assay was used to detect the proliferation ability. Results Compared with control group and mutation group, the expressions of miR-200a/b/c were significantly de-creased, especially for miR-200b. And the expressions of ZEB1/ZEB2 were significantly increased at both mRNA and pro-tein levels after transfected with the ZEB2 3′UTR, enhancing the capability of migration,invasion,and proliferation (P <0.05). Compared with ZEB2 3′UTR group, the capabilities of proliferation,invasion and migration were significantly lower in combined group. Conclusion ZEB2 3′UTR can increase the ability of cell proliferation, invasion and metastasis through regulating the levels of miR-200a/b/c, and then influence the regulation of transcription of the target gene, which could lead to malignant transformation of GES-1 cells.

Objective To explore the curative effect of knee flexion deformity on spastic cerebral palsy treatment method.Methods All of 30 patients with spastic cerebral and knee flexion deformity were randomly divided into two groups:traditional operation group and modified operation group,using the commonly used operation (In 15 cases,with traditional hamstring post surgery) and) modified operation (In 15 cases,with modified hamstring post surgery),two groups of patients were treated with Ilizarov external fixation drafting device in the correction of knee flexion deformity after soft tissue release.Adjustment began 7 days after the soft tissue release operation of external fixation,and stretched approximately 0.2 degrees each time,3 times/d,until knee flexion deformity was corrected to hyperextension for 10 degree and maintained for 3 weeks,and the flexion contracture degree of knee joint was measured every 2 weeks;then the Ilizarov external fixation drafting device was removed.Then wear a walking straight leg brace for more than 3 months,until the knee walking function is good.Clinical evaluation included the joint pain index,walking function index,knee flexion deformity degree and range of joint motion score of Dimeglio standard.Results Flexion contracture deformity of knee joint in 30 cases (60 knees) was corrected when Ilizarov external fixator was removed,knee extension to 0 degrees-5 degrees.The two groups of patients had the knee flexion angle range of-1.2 degrees to 13.3 degrees,with an average of (7.32°±3.41°) after removed of the walking straight leg brace,in which 4 cases (8 joints) got recurrent deformity of 10°-15° at the time of removing of the walking straight leg brace.Knee activity significantly was improved at the end of treatment.60 cases of knee joint activity were close to normal,with flexion of 100 degrees to 135 degrees,extension of 0 degrees to 10 degrees.Two groups of patients were statistically significantly improved before and after surgery.Curative effect comparison:The walking function index of the modified operation group was obviously superior to that of the traditional operation group,there was significant statistical difference.There were no statistically significant differences in pain index,knee flexion,and range of joint motion.Conclusion For the treatment of flexion deformity of the knee joint in spastic cerebral palsy,traditional surgery using the semitendinosus and gracilis,post and semimembranosus lysis,combined with the Ilizarov draft external fixation could improve the walking function of the patients,simplify the surgical incision and reduce trauma.As a result,modified hamstring post surgery is an ideal,effective treatment method.

Objective:To show whether the Fas Ligand gene induces mast cells apoptosis.Methods:RT-PCR was used to amplify the gene of rat Fas ligand extracelluar domain and transmembrane domain and cloned it into eukaryotic expression plasmid pcDNA3.1.Transcent transfect RBL-2H3,the expression of Fas ligand RBL-2H3 was detected by RT-PCR、Western blot.The Annexin V FCM was used to detect the RBL-2H3 apoptosis after the transfection of Fas Ligand.Results:It is successful to obtain the gene of rat Fas Ligand extracellular domain and transmembrane segment,cloning it into pcDNA3.1,FasL was expressed on the surface of RBL-2H3 and it's supernatant after the transfection of pcDNA3.1/FasL.The cell start to be apoptosis.Conclusion:Our study reveals that Fas Ligand gene transfection in RBL-2H3 can effectively induced apoptosis.It is a promising strategy for Fas Ligand to be used in the therapy of allergic disease. [

OBJECTIVE: To explore the mechanism of Flos Puerariae and Semen Hoveniae in treatment of alcoholic liver injury (ALI) based on network pharmacology and molecular docking. METHODS: The information of chemical constituents and targets of Flos Puerariae and Semen Hoveniae was collected from TCMSP and Swiss databases, and the threshold values of oral bioavailability (OB) ≥ 30%, drug likeness (DL) ≥0.18 were used to screen the potential active compounds. The GeneCard and DrugBank databases were used to obtain the targets corresponding to ALI. The common targets were queried using Venn Diagram, and the network of PPI and Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed through DAVID and Reactome database. Autodock Vina software was used for molecular docking of potential ingredients and key targets. RESULTS: A total of 21 potential active compounds and 431 therapeutic targets were gathered in Flos Puerariae and Semen Hoveniae, which involved 273 biological functions, 90 KEGG pathways and 362 Reactome pathways. The GO functions involved protein binding, ATP binding, etc.; the KEGG pathways mainly included PI3K-Akt signaling pathway and TNF signaling pathway; the Reactome pathways contained signal transduction and immune system, etc. The results of molecular docking showed that 21 potential active ingredients had good affinity with the core targets Akt1, TP53 and IL-6. CONCLUSIONS The network pharmacology and molecular docking analysis demonstrate the synergetic effect of Flos Puerariae and Semen Hoveniae with multi-compounds, multi-targets and multi-pathways in the treatment of ALI; and also predict the possible medicinal substance, key targets and pathways, which provides clues for the new drug development and mechanism research.
Animals ; Computer Simulation ; Drugs, Chinese Herbal/therapeutic use* ; Lepidoptera/chemistry* ; Liver/drug effects* ; Liver Diseases, Alcoholic/therapy* ; Molecular Docking Simulation ; Phosphatidylinositol 3-Kinases/metabolism* ; Plant Extracts/therapeutic use* ; Rhamnaceae/chemistry* ; Signal Transduction/drug effects*

Medical device imaging data augmentation is a method of expanding existing datasets by generating new data samples,which is of great significance for improving the performance of artificial intelligence(AI)medical device-related models and clinical application effects.However,traditional data augmentation methods are usually limited by the quality,realism,and diversity of generated samples.Denoising diffusion probabilistic model(DDPM)is a generative model based on the noise diffusion process,and its main idea is to generate samples with high quality by modelling the sampling process of the target distribution as a process of progressive denoising from the noise distribution.The basic principles and working mechanisms of DDPM were reviewed,the application scenarios of this method in AI medical device data augmentation were analyzed,and its advantages,challenges,and future development directions were explored to provide a reference for the field of AI medical device data augmentation.