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OBJECTIVE@#To analyze the subcellular localization of GTPase of immunity-associated protein 2 (GIMAP2) for the further functional study.@*METHODS@#In the study, we first obtained the protein sequences of GTPase of immunity-associated protein 2 (GIMAP2) from National Center for Biotechnology Information (NCBI) database, and then performed a prediction analysis of its transmembrane structure, nuclear localization signal (NLS), nuclear export signal (NES) and subcellular localization through bioinformatics online tools. GIMAP2 gene amplified by PCR was inserted into the expression vector pQCXIP-mCherry-N1 and positive clones were selected by ampicillin resistance. After using methods to extract and purify, the sequenced recombinant plasmid pQCXIP-GIMAP2-mCherry, together with the retroviral packaging plasmids VSVG and Gag/pol, was transferred into HEK293FT cells by liposomes for virus packaging. The virus supernatant was collected 48 h after transfection and directly infected the human breast cancer cell line MDA-MB-436. Immunofluorescence staining was constructed to detect the localization of endogenous and exogenous GIMAP2 in MDA-MB-436 cells. Meanwhile, green fluorescent chemical dyes were used to label mitochondria, endoplasmic reticulum, and lipid droplets in living MDA-MB-436 cells stably expressing the GIMAP2-mCherry fusion protein. Images for the three dye-labeled organelles and GIMAP2-mCherry fusion protein were captured by super-resolution microscope N-SIM.@*RESULTS@#Bioinformatics analysis data showed that GIMAP2 protein composed of 337 amino acids might contain two transmembrane helix (TM) structures at the carboxyl terminus, of which TMs were estimated to contain 40-41 expected amino acids, followed by the residual protein structures toward the cytoplasmic side. NES was located at the 279-281 amino acids of the carboxyl terminus whereas NLS was not found. GIMAP2 might locate in the lumen of the endoplasmic reticulum. Sequencing results indicated that the expression vector pQCXIP-GIMAP2-mCherry was successfully constructed. Fluorescent staining confirmed that GIMAP2-mCherry fusion protein, co-localized well with endogenous GIMAP2, expressed successfully in the endoplasmic reticulum and on the surface of lipid droplets in MDA-MB-436 cells.@*CONCLUSION@#GIMAP2 localizes in the endoplasmic reticulum and on the surface of LDs, suggesting potential involvement of GIMAP2 in lipid metabolism.
Subject(s)
Humans , Amino Acid Sequence , Cytoplasm , GTP Phosphohydrolases , Membrane Proteins , Nuclear Export Signals , Nuclear Localization Signals , Recombinant Fusion Proteins , TransfectionABSTRACT
Abstract?AIM:To evaluate the effect of combined glaucoma and cataract surgery on the function of eye surface and tear film.?METHODS:This clinical trial involved 75 patients ( 75 eyes ) with glaucoma complicated with cataract undergoing combined glaucoma and cataract surgery.All the eyes were divided into surgical group ( the eyes undergoing operation ) and control group ( the other eyes).The symptoms and signs of dry eye disease, Schirmer's I test, break -up time ( BUT ) , corneal fluorescein staining were tested and marked 1mo after operation.Comparison of the results was made between two groups.?RESULTS:The average intraocular pressure was 16.25± 0.46mmHg at 1mo after operation, compared with 45.29± 4.39mmHg at 3d before surgery, the difference was statistically significant ( P<0.05 ).Preoperative average visual acuity was 0.08±0.06, corrected visual acuity was<0.05 in 23 eyes,≥0.05 to<0.1 in 16 eyes,≥0.1 to<0.3 in 36 eyes.Postoperatively 1mo, corrected visual acuity:<0. 05 for 7 eyes (9%),≥0.05 to <0.1 for 11 eyes (15%), ≥0.1 to<0.3 for 38 eyes (51%), ≥0.3 to <0.5 for 11 eyes (15%) , ≥0.5 for 8 eyes (11%), postoperative visual acuity was 0.15 ±0.1, which was significantly higher than before surgeries ( P<0.05).The postoperative dry eye symptoms of surgical group was higher, compared with that of control group, and the differences of diagnosis of dry eye test indicators between the two groups were statistically significant(P<0.05).? CONCLUSION: Combined cataract and glaucoma surgery may affect postoperative ocular surface and tear film function, make the tear film stability damaged and lead to dry eye disease.
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<p><b>OBJECTIVE</b>To study clinical effects of plaster combined with splint for the treatment of Gartland type 1I humeral fractures.</p><p><b>METHODS</b>From March 2002 to May 2006, 24 children with humeral supracondylar fractures of Gartland type ill were reviewed. Among the patients, 14 patients were male and 10 patients were female, ranging in age from 4 to 12 years, averaged 6.6 years. Ten patients had injuries in the left and other 14 patients had injuries in the right limb. Firstly, the patients were treated with manipulative reduction to maintain the length of humerus without emphasis on anatomic reduction. Then, the patients were treated with external fixation using plaster for 5 to 7 days, and secondary manipulative reduction after swelling disappeared. Lastly, the patients were treated with external fixation using splint for 4 to 5 weeks until fracture union.</p><p><b>RESULTS</b>All the patients were followed up, and the duration ranged from 5 months to 2 years, with an average of 1.2 years. All the patients had no complications such as neurovascular injury, myositis ossificans, forearm compartment syndrome and Volkmann contracture. According to ZHU Xiao-ting evaluation criteria for humeral supracondylar fractures in children, 12 patients got an excellent result, 8 good, 3 poor and 1 bad.</p><p><b>CONCLUSION</b>Treatment of child humeral supracondylar fractures with plaster and splint has several advantages such as avoiding serious soft tissue injuries around fractures due to many times reduction, fracture dynamic correction to obtain satisfactory reduction, reducing complications, obtaining good reduction, and restoring elbow function in a relatively short period of time.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Male , Casts, Surgical , Humeral Fractures , Diagnostic Imaging , General Surgery , Radiography , SplintsABSTRACT
Objective To determine the value of Magnetic resonance imaging(MRI)technique on diagnosis of adrenal medulla hyperplasia model rats confirmed by pathology study.Methods Sixty male SD rats were divided into 6 groups randomly,10 rats in each group.As experiment group,A,B,C groups were subcutaneously injected with reserpine(0.4 mg?kg~(-1)?d~(-1))from the beginning to 40,60,80 days respectively.As compare,the control groups(a,b,c group)were only injected with sodium chloride simultaneously.MRI technique and pathology study were performed for all the subjects on 40,60,and 80 days respectively.Results The percent of medulla were higher in B and C groups than that in b and c groups respectively[(34.3?5.8)% vs.(25.7?8.9)%,t=2.462,P