Objective To observe the effect of subretinal injection of retinal pigment epithelium (RPE) cells for RPE in mice.Methods A total of 30 postnatal day 7 C57BL/6J mice were randomly divided into normal mice group,OIR model group and OIR model cell transplanted group,10 mice in each group.The OIR model was induced in mice of OIR model group and OIR model cell transplanted group.The RPE cells were subretinal injected into the RPE of mice in OIR model cell transplanted group.At 20 days after the injection,the RPE thickness was evaluated by fluorescence microscope.The expression of RPE65,Bestrophin and zonula occludens-1 (ZO-1) were estimated by Western blot and real-time quantitative PCR (RT-PCR).Results The thickness of RPE in OIR model mice was thinner than that in normal mice;the thickness of RPE in OIR model cell transplantation mice was significantly thicker than that in the OIR model mice.The results of Western blot and RT-PCR indicated that the differences of protein (F=8.597,18.864,25.691) and mRNA expression (F=39.458,11.461,34.796) of RPE65,Bestrophin,ZO-1 were statistically significant between OIR model group and OIR model cell transplanted group (P＜0.05).Conclusions Subretinal injection of RPE cells can promote RPE thickening.RPE65 and Bestrophin protein relative expression levels increased,ZO-1 protein relative expression levels reduced;mRNA expression levels of RPE65,Bestrophin and ZO-1 genes increased.

Burns ; complications ; immunology ; physiopathology ; Humans ; Hypoxia ; etiology ; immunology ; physiopathology ; Ischemia ; etiology ; immunology ; physiopathology

AIM: To investigate the effects of microRNA (miRNA)-483-3p on the growth and migration of human glioma cell line A172 and its potential mechanisms.METHODS: The abundance of miRNA-483-3p in human embryonic kidney 293 cells and different human glioma cell lines (A172, U251 and SHG44) was measured by RT-qPCR.After down-regulation of miRNA-483-3p by transfection of inhibitor in the A172 cells, the cell viability, cell cycle distribution and cell migration were detected by CCK-8 assay, flow cytometry and Transwell assay, respectively.Furthermore, the protein levels of cell cycle-related molecules and epithelial-mesenchymal transition markers were measured by Western blot.Luciferase reporter assay was used to predict and verify the target gene of miRNA-483-3p.RESULTS: miRNA-483-3p was highly expressed in human glioma cells.Knockdown of miRNA-483-3p inhibited A172 cell viability, arrested cell cycle and decreased cell migration rate.Furthermore, the protein levels of cyclin D1, cyclin-dependent kinase 4, phoshorylated retinoblastoma protein, N-cadherin and vimentin were significantly decreased after knockdown of miRNA-483-3p, accompanied with the up-regulation of E-cadherin and β-catenin protein expression.Luciferase reporter assay demonstrated that Smad4 was a potential target gene of miRNA-483-3p.Down-regulation of Smad4 in the A172 cells transfected with miRNA-483-3p inhibitor partially reversed the effect of miRNA-483-3p on cell viability and migration.CONCLUSION: Knockdown of miRNA-483-3p restrains the growth and migration of A172 cells by targeting Smad4.

Chronic Disease ; therapy ; Constipation ; diagnosis ; therapy ; Gastrointestinal Transit ; Humans

According to the synonymous codons used in 28 open reading frames from Pichia pastoris, the codon usage in this species was calculated and 19 codons have been inferred to be its optimal codons. The results show that pattern of the codon usage in P. pastoris is similar to that in S. cerevisiae and in K. lactis except for the synonymous codon of glutamic acid, which may be the special bias of P. pastoris.

Objective To analyze and evaluate the safety and efficacy of modified Pie-crusting ( PC) technique for releasing medial tightness during primary total knee arthroplasty ( TKA) . Methods We completed primary TKA by the same performer with modified PC technique in 30 patients (34 knees) with genu varus from March 2014 to June 2016.By using a special curved scalpel with width limit of 3 mm and depth limit of 5 mm to poke the tension parts , we released the anterion bondle of superficial medial collateral ligament ( sMCL) and posteromedial corner structures ( PMCS) during tension happened in extension , and we released the posterior bondle of sMCL during tension happened in flexion .According to the gap value measured intraoperatively , we divided these cases into three groups: extension with flexion tension group ( 10 knees ) , extension tension group ( 13 knees ) , and flexion tension group (11 knees).The difference between medial and lateral gap value no more than 1 mm was defined as gap balance.We calculated the gap balance rate of each group .Series of weighted frontal X-ray were conducted at fixed period to evaluate the varus angle of the knee postoperatively.The range of motion (ROM), HSS scores and WOMAC scores were also recorded at the same time . Results Among the 34 knees , 31 knees reached the medial and lateral gap balance at both extension and flexion .There was a difference of 2 mm in medial and lateral gap value at extension in 1 knee and the same difference at flexion in the other 2 knees.The total postoperative gap balance rate was 91.2%( 31/34 ).The constrained inserts were implanted in 3 cases.No technical-related complications happened after the surgery.After the releasing procedure, the flexion gap value had an increase of 1 mm (range, 1-3mm) in the extension tension group , and the extension gap value had an increase of 1 mm ( range, 1-2 mm) in the flexion tension group, without significant difference (Z=-1.118, P=0.264).The ROM was 83.3°±14.7°preoperatively and 100.7°±14.2° postoperatively (t=-7.714, P=0.000).The median alignment of the knee was 11.5°(range, 7°-32°) preoperatively and 1° (range, 0°-4°) postoperatively (Z=-5.092,P=0.000).The HSS scores were (42.7 ±16.3) points preoperatively and (88.1 ± 9.9) points postoperatively (t=-21.868, P=0.000).The WOMAC scores were (76.2 ±8.2) points preoperatively and (11.4 ± 9.7) points postoperatively (t=31.726, P=0.000).All of them were significantly improved in comparison with those before the surgery. Conclusions Using modified PC technique is safe and effective in medial releasing during primary TKA .Both extension and flexion gap value will be affected by releasing tensed fiber at extension or flexion position .

OBJECTIVEThis research aimed to fabricate a hydroxyapatite (HA)-chitosan scaffold via simulated body fluid (SBF) biomimetic mineralization and determine how mineralization time affects scaffold construction and cell compatibility.

METHODSThe HA-chitosan scaffolds were fabricated by freeze-drying technique and then subjected to precalcification, also known as alternative soaking method. Afterward, precalcificated scaffolds were placed into the SBF to conduct the mineralization process. Mineralization time was set at 7, 14, and 21 days, corresponding to three experimental groups. Pure chitosan scaffolds acted as the control group, and the physical and chemical properties of the four groups were tested. Osteogenic-induced adipose-derived stem cells (ADSCs) were seeded into the scaffolds to investigate the scaffolds' cell compatibility.

RESULTSThe mineral substance of the 14-day group exhibited a uniform distribution. The crystal composition of the mineral substance suited the HA's features. The compressive elastic modulus increased along with the extension of mineralization time. The 21-day group showed a statistically significant increase in compressive elastic modulus compared with the control group (P < 0.05). The cell proliferation level of the 14-day group was significantly the highest among the three experimental groups (P < 0.05). The calcium ion and the type I collagen had the highest secretion amount when the cells were seeded into the 14-day group.

CONCLUSIONThe SBF biomimetic mineralization method can be used to fabricate HA-chitosan bone-tissue-engineering scaffolds. The biological compatibility, as well as the chemical and physical properties, reached the optimum levels at day 14.

Purpose To study X-ray and CT findings of pulmonary infection in AIDS. Methods We evaluate 10 cases who have chest abnormalities. The all patients have X-ray and CT examination. Results Imaging of pulmonary tuberculosis in AIDS, including infiltrative imaging and diffuse patchy imaging, enlarged mediastinum lymph nodes and extra chest enlarged lymph nodes. The imaging of legionella species is infiltrative imaging. Carinii pneumonia appearances diffuse infiltrative and interstitial abnormalities. Conclusion Pulmonary tuberculosis, legionella species and carinii pneumonia are common infections in AIDS. The patchy, consolidations, diffused diseases and enlarged lymph nodes are important imaging appearances.

Objective To investigate the clinical and imaging features of cerebral cortical vein thrombosis (CoVT). Methods The clinical data of 13 inpatients with CoVT were enroled from September 2010 to September 2015. The etiology, clinical symptoms, imaging features, and differential diagnosis of CoVT w ere summarized and analyzed. Results Among the patients, there w ere 7 females (53.85%) and 6 males (46.15%). Their age ranged from 18 to 59 years (mean 34.23 ±12.08). The most common clinical manifestations were headache (9 patients) and seizures (9 patients). The 13 patients had cerebral parenchymal involvement and they al received anticoagulant therapy. Conclusions The clinical symptoms of CoVT are various. Seizures and headache are the most common symptoms. Most of patients are accompanied by brain parenchymal lesions and venous sinus thrombosis. When patients have headache and seizures, and head images suggest hemorrhagic infarction, imaging examination of the improvement of the cerebral venous system thrombosis is recommended.

Blood Gas Analysis ; Female ; Humans ; Infant, Newborn ; Infant, Newborn, Diseases ; therapy ; Intensive Care Units, Neonatal ; Male ; Respiration, Artificial