Objective For the purpose of exploring the influence exerued by the amount of template DNA on the results of automatic fluerescent detection. Method For the purpose of buiding the optimal amount of template DNA amplified, we have analyzed different amount of template DNA (9947A) by 3100 Genetic Analyzer with the multiplexing Profiler Plus Kit. Results The optimal amounts of template DNA amplified were between 0.31ng and 2. 5ng by using 3100 Genetic Analyzer. Conclusion It will produce the incorrect analytic results if the amounts of template DNA were too high or too low.

Child, Preschool ; Fever ; etiology ; Histoplasmosis ; complications ; diagnosis ; pathology ; Humans ; Jaundice ; etiology ; Male

imaging features of myositis ossificans have some characteristics. Misdiaguosis could be avoided when the disease was evaluated with the course.

Objective:To investigate the effect of miRNA-5089-5p (miR-5089-5p) on proliferation and migration ability of esophageal cancer in vitro and its relationship with the expression of cathepsin B (CTSB) gene.Methods:Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-5089-5p in 31 tissue samples from patients who underwent esophageal cancer resection and the corresponding pericarcinomatous tissues between in March 2017 and in December 2019 at Huangshi Central Hospital of Edong Healthcare Group, and TE-13, EC9706, Eca109, KYSE30 cell lines and normal esophageal mucosal epithelial HET-1A cells. The esophageal cancer cells with the lowest expression level of miR-5089-5p were divided into 2 groups: miR-5089-5p group transfected with miR-5089-5p mimics and the negative control group with negative control sequence. qRT-PCR was used to detect the expression level of miR-5089-5p after transfection for 48 h. CCK-8 method and scratch healing test were used to detect the proliferation and migration ability of cells in the two groups. The online tools microRNA.org and Deepbase v2.0 were used to predict the target genes of miR-5089-5p. The dual luciferase reporter gene assay was used to verify the target gene of miR-5089-5p. qRT-PCR and Western blot were used to detect the expression level of target genes in the two groups. The expressions of cell proliferation-related protein (PCNA and Ki-67) and migration-related protein (N-Cadherin and Twist) were detected by using Western blot.Results:The relative expression level of miR-5089-5p in esophageal cancer and pericarcinomatous tissues was 1.54±0.53 and 7.07±1.25, respectively ( t = 24.06, P < 0.01). The relative expression level of miR-5089-5p in the esophageal cancer cell lines was lower than that of normal esophageal mucosal epithelial HET-1A cells (all P < 0.05), and the cell line with the lowest relative expression was Eca109 cells (0.12±0.03). Compared with the negative control group, the proliferation ability of Eca109 cells in miR-5089-5p group was gradually reduced with the transfection time extension, and the difference was statistically significant between the two groups since 48 h (all P < 0.05), and the migration ability was also reduced [scratch healing rate: (29±5)% vs.(64±8)%, t=3.91, P < 0.01]. The online tool predicted that the target gene of miR-5089-5p might be CTSB, and the dual luciferase reporter gene assay confirmed that miR-5089-5p complemented CTSB 3'UTR. qRT-PCR results showed that compared with the negative control group, the relative expression level of CTSB mRNA in Eca109 cells of miR-5089-5p group was reduced (0.23±0.04 vs.1.01±0.09, t = 8.27, P < 0.01). Western blot results showed that the expression level of CTSB protein was reduced, and the expression levels of cell proliferation-related protein PCNA, Ki-67 and cell migration-related protein N-Cadherin, Twist were also reduced. Conclusions:The expression level of miR-5089-5p in esophageal cancer tissues and cell lines is low. miR-5089-5p can inhibit proliferation and migration of esophageal cancer Eca109 cells. The mechanism may be achieved by down-regulating CTSB gene expression.

Vitexin, a naturally occurring flavone glycoside in plants, has many pharmacological effects, which is widely distributed in nature. This paper reviewed the research progress of the distribution of vitexin in the plant resources and its pharmacological effects, and summarized its application prospects, aiming to provide a useful reference for the development of vitexin-enriched plant resources.
Animals ; Antineoplastic Agents ; pharmacology ; Antioxidants ; pharmacology ; Apigenin ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Myocardial Infarction ; drug therapy ; Plant Dispersal

Anion Exchange Protein 1, Erythrocyte ; metabolism ; CD56 Antigen ; metabolism ; Desmin ; metabolism ; Desmoplastic Small Round Cell Tumor ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Humans ; Lymphoma ; metabolism ; pathology ; Male ; Middle Aged ; Rhabdomyosarcoma ; metabolism ; pathology ; Sarcoma, Ewing ; metabolism ; pathology ; Testicular Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.
Animals ; Base Sequence ; Cloning, Organism ; Electrophoresis ; Endonucleases ; genetics ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Gene Knockout Techniques ; Gene Targeting ; methods ; Goats ; Green Fluorescent Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Zinc Fingers

Objective To explore the clinical value of guide-wire shaping in subclavian vein catheter-ization.Methods Totally 400 patients requiring right subclavian vein catheterization were equally divided into two groups according to the clinic date: intervention group ( with guide-wire shaping , n =200 ) and control group (without guide-wire shaping, n=200).The catheterization was carried out by the same doctor .The rates of ectopic wire were compared between the two groups .Results The overall success rate of catheteriza-tion was 98.25%(393/400) [98.5% (197/200) in intervention group and 98.0% (196/200) in control group, P=0.500].The incidence of catheter displacement was 1.02%(2/197) in intervention group, which was significantly lower than that [7.14% (14/196)] in control group (P=0.002).Conclusion As a sim-ple procedure , guide-wire shaping can effectively prevent catheter displacement during catheterization .

Background and purpose:Positron emission tomography-computed tomography (PET/CT) is playing an increasingly important role in the diagnosis, therapy and follow-up of lymphoma patients. This study aimed to explore clinical and pathological features and bone marrow infiltration status in lymphoma patients with diffused high bone marrow glucose uptake on18F-FDG PET/CT.Methods:It was a retrospective study. Bone marrow infiltration status, pathological and clinical data from 62 cases of pathologically diagnosed lymphoma and diffused high bone marrow glucose uptake were analyzed.Results:Distribution of histopathological subtype in those cases was in accordance with that in previously reported Chinese lymphoma patients. Significant difference was demonstrated in standard uptake value (SUV) between pa-tients with aggressive and indolent histopathological subtypes (8.43vs 5.38,P=0.048), patients with and without B symp-toms (8.30vs 5.72,P=0.033), and patients with and without bone marrow infiltration (8.78vs 6.96,P=0.020). 32 patients were diagnosed as “bone marrow infiltration” by bone marrow biopsy. There was significant difference in histopathologi-cal subtype distribution between patients with and without bone marrow infiltration (P=0.001). In patients with bone marrow infiltration, there were higher proportions of mantle cell lymphoma, nodal marginal zone B cell lymphoma, Burkitt’s lym-phoma and anaplastic large cell lymphoma. In contrast, patients without bone marrow infiltration suffered more from diffuse large B-cell lymphoma, peripheral T cell lymphoma, enteropathic T cell lymphoma and extranodal NK/T-cell lymphoma (nasal type). False positive results in bone marrow glucose uptake may be caused by fever or anemia.Conclusion:Diffused high bone marrow glucose uptake on18F-FDG PET/CT should be evaluated in combination with the uptake values, clinical features and histological subtypes, to minimize the misdiagnosis and to better guide staging and therapy of lymphoma.

Objective To evaluate the feasibility and safety of embedding the totally implantable venous access port (TIVAP) via the access of right brachiocephalic vein (BCV).Methods The clinical data of 493 patients,who underwent the placement of TIVAP by using right BCV route during the period from March 2013 to December 2015,were retrospectively analyzed.The patients included 137 males and 356 females,with a mean age of (47.3±13.2) years old (ranging from 29 to 78 years old).The puncture success rate and TIVAP indwelling procedure-related complications were analyzed.Results The technical success rate was 100％,the success rate of initial puncturing was 99％ (488/493).The mean operation time was (22.5± 8.3) minutes (range of 18-35 minutes).Mis-puncturing of artery happened in 3 patients (0.61％,3/493);and no severe complications such as hemothorax or pneumothorax occurred.After implantation,the patients carried TIVAP for 124-986 days,with a mean of (271.1±53.8) days.The incidence of complications was 2.25％ (11/488),including hemorrhage at port site (n=2),catheter-related infection (n=l),partial thrombosis (n=2),and formation of fibrous protein sheath (n=6).No serious complications such as displacement or rupture of catheter,or catheter pinch-off syndrome (POS),etc.were observed.Conclusion The implantation of TIVAP by using right BCV route has high puncturing success rate,the technique is safe and reliable,and it can provide another option of catheter access for the clinical performance of TIVAP implantation.