Purpose To study the effect and mechanism of miR-199a-5p on the invasion of breast cancer MDA-MB-231 cells. Meth-ods miR-199a-5p mimic was transfected into MDA-MB-231 cells. Influence of miR-199a-5p on the invasion of MDA-MB-231 cell was displayed by Transwell, the expression of epithelial-mesenchymal transition ( EMT) molecular markers ( E-cadherin, vimentin) regulated by miR-199a-5p was determined using immunofluorescence and Western blot. Western blot was employed to assess the levels of ERK5, pERK5, EGF and SP1 in MDA-MB-231 cells dealt with miR-199a-5p mimic and LNA-siRNA. Chromatin immunoprecipita-tion (CHIP) was applied for displaying the reaction of SP1 with ERK5 promoter. Results miR-199a-5p could inhibit the invasion of MDA-MB-231 cells, decrease the expression of vimentin and enhance E-cadherin. Meanwhile, miR-199a-5p decreased the expression of ERK5 and pERK5, the levels of EGF and SP1 were also downregulated. On the contrary, the levels of EGF, SP1, ERK5 and pERK5 were enhanced by employing LNA-siRNA targeting miR-199a-5p. SP1 could bind with ERK5 promoter. Conclusions miR-199a-5p could reduce the expression of ERK5 and pERK5 through regulating EGF and SP1, which functioning the inhibitory effect on invasion of MDA-MB-231 breast cancer cells.

Objective To observe the long term results of intertrochanteric varus medial displacement osteotomy(IVMDO) for Perthes disease. Methods Thirty eight patients with Perthes disease treated with IVMDO were reviewed. The results were evaluated based on a criteria made by the authors including clinical and radiographic parameters. The duration of follow up ranged from 3 to 15 years, with an average of 7 years. Results Fifteen patients were evaluated as having excellent result, 17 good, 3 fair and 3 poor respectively. The overall excellent or good rate was 84.2%. Considering the relationship between the outcome and staging of the disease, the overall excellent or good rate was 94.7% in stage Ⅱ lesion, 85.7%in stage Ⅲ lesion, and 40.0% in stage Ⅳ lesion. Conclusion The treatment of Perthes disease with IVMDO has the advantages of simple manipulation, less trauma and good results, and is worthy of populariztion.

Objective To investigate clinical and pathological features of gastrointestinal stromal tumors,the frequency and type of mutation of c-kit and platelet-derived growth factor receptor α (PDGFRA)genes.Methods 340 patients underwent surgical resection and diagnosed as gastrointestinal stromal tumors by postoperative pathology from Junuary 2012 to December 2014 were enrolled,and their tumor tissues were collected.The direct sequencing method was applied to detect the mutation status of c-kit gene (exon 9,11,13 and 17) and PDGFRA gene (exon12 and 18).Results There were 138 males and 192 females,and their median age was 58 years old (37-81 years old).There were 178 patients (52.4 ％) with gastric stromal tumors,21 cases (6.2 ％) with duodenal stromal tumors,82 cases (24.1％) with small intestinal stromal tumor,10 cases (2.9 ％) with colon stromal tumor,15 cases (4.4 ％) with rectal stromal tumors,30 cases (8.8 ％) with parenteral stromal tumor (from the peritoneum,mesentery,retroperitoneum or attachment),4 cases (1.2 ％) of liver tissues (gene detection tissues from the liver biopsy or surgical resection specimens).In the mutation analysis of all 340 patients with gastrointestinal stromal tumor,the total mutation rate was 89.4 ％ (304/340),including 81.2 ％ (276 cases) of c-kit,8.2 ％ (28 cases) of PDGFR and 10.6 ％ (36 cases) of wild type.Among 125 cases underwent the detection of the gene mutation sequences,there were 49 cases of exon11 deletion mutation,4 cases of exon11 insertion mutation,12 cases of exon 11 missense mutation,8 cases of exon9 insertion mutation,1 case of 3xon13 missense mutation,1 case of exon17 missense mutation,24 cases of exon12 synonymous mutation and 20 cases of exon18 synonymous mutation.Conclusions Gene detection is becoming more and more obvious in predicting the therapeutic effect of molecular targeted therapy,the mechanism of drag resistance and the clinical treatment,c-kit exon11 mutation is one of the most common gene mutation types related to the choice of targeted medicine.

OBJECTIVE To investigate the short-term clinical efficacy of sublingual immunotherapy in children with allergic rhinitis. METHODS Forty-four patients, from 3-14 years of age, with mite-sensitive allergic rhinitis were treated sublingually with standardized dermatophagoidesfarinae extract. After treatment of 3 or 6 months using the dermatophagoidesfarinae extract, the total nasal symptoms score(including nasal congestion, nasal itching, rhinorrhea and sneezing scores), total rhinitis medicine scores, visual analog scale scores and the percentage of blood eosinophilic granulocyte(Eos%) before and after treatment were evaluated. SPSS 20.0 software was used to analyze the data. RESULTS After 3 or 6 months' treatment,thenasal symptoms score(including nasal congestion, nasal itching, rhinorrhea and sneezing scores), total rhinitis medicine scores and visual analog scale scoreshad significantly reduced,comparing with that before treatment(all P<0.05). The comparison of the total nasal symptoms score(including nasal congestion, rhinorrhea and sneezing scores), total rhinitis medicine scores and visual analog scale scores between 3 months and 6 months treatment had shown no significant difference (all P>0.05), while nasal itching scores had significantly reduced(Z=-4.185, P<0.05). After 6 months' treatment, the percentage of Eos% had significantly reduced compared with that before treatment(Z=-4.544, P<0.05). CONCLUSION SLIT could be an effective treatment in patients with allergic rhinitis sensitive to house dust mite. It would have effect after3 months treatment and Eos%reduced significantly after 6 months treatment.

OBJECTIVE To discuss the diagnosis and endoscopic treatment of congenital basal encephaloceles in infants.METHODS We retrospectively reviewed the clinical data of 6 infants with congenital basal encephaloceles in Beijing Children's Hospital between January 2014 to September 2016. CT and MRI were performed routinely. All patients underwent endoscopic resection of encephaloceles and repair of skull base defects. RESULTS Five patients presented with transethmoidal encephaloceles and one transsphenoidal encephaloceles. All patients underwent endoscopic procedure successfully. There were no complications except for one 7-month old girl who got purulent meningitis. All patients had favorable clinical outcomes during a follow-up of 3 to 29 months.CONCLUSION For infants with persistent nasal obstruction and nasal neoplasms, congenital basal encephaloceles should be considered. Nasal coronary CT and sagital MRI are of paramount importance in the diagnosis of congenital basal encephaloceles in infants. The endoscopic procedure is the safe and effective method for the management of congenital basal encephaloceles in infants.

Purpose To investigate the expressions of PKCζ, MMP-2, and MMP-9 in breast cancer and the relationship with the inva-sion and metastasis of breast cancer. Methods The expression of PKCζ, MMP-2 and MMP-9 in 100 cases with breast cancer was as-sessed with immunohistochemistry PV 9000 method. PKCζ-siRNA was transfected into MDA-MB-231 cell lines, called siPKCζ/MDA-MB-231. While siRNA construct containing a scrambled sequence was transfected into MDA-MB-231 cells to generate control cells, which were designated as Scr/MDA231 cells. Western blotting was used to measure the expression of PKCζ in transfected cells, and the Transwell invasion assay was used to detect the invasion ability in vitro. The content of MMP-2, MMP-9 were measured by ELISA. Results The expression rates of PKCζ, MMP-2 and MMP-9 in breast cancer tissues were 62.5%, 37.5% and 32.5%, and there were significant differences among them (P<0.05). The expression of PKCζwas much higher than those in the normal breast tissues nearby. The expression of PKC protein was assoiated with lymph node metastasis, distant metastasis (P<0.01), but was not correla-ted with other clinicopathologic parameters, such as age, tumor size, histological type, ER, PR, and so on (P>0.05). The expres-sion of PKCζ, MMP-2 and MMP-9 were lower in siPKCζ/ MDA-MB-231 group than those in scr/ MDA-MB-231 group, and the in vitro invasion ability was significantly decreased (P<0.05). Conclusions PKCζ can promote the invasion and metastasis of breast canc-er, and correlated with the expression of MMP-2, MMP-9(P<0.05).

Objective:To study the effect of sublingual immunotherapy on children with mite allergic rhinitis.Methods:Four hundred and ninety patients with mite allergic rhinitis have been recruited by Beijing Children′s Hospital from March 2014 to September 2019 and divided into 4 groups of young children group, different treatment duration group, individualized dose adjustment group and multiple allergy evaluation group, the clinical scores of total nasal symptoms score (TNSS), visual analogue scale scores (VAS) and total medication score were recorded at the first visit, 12 months, 24 months and 36 months after treatment, and the combined symptom and medication score(CSMS) score was calculated.Results:A total of 374 patients (76.32%) completed this study.Among them, the CSMS(2.20±1.61, 2.50±1.78), TNSS(2.80±2.32, 3.60±2.71) and VAS(3.50±1.16, 3.90±1.43) in ≤3-year-old group and children over 3-year-old group of young children set after use of 12 months were significantly lower than the score at the first time of diagnosis (respectively CSMS: 4.50±1.44, 5.30±1.32; TNSS: 6.20±1.89, 7.50±2.19; VAS: 5.40±2.33, 5.90±1.61). In addition, in the duration and efficacy set, the patients who completed the immunotherapy for 36 months can only be observed in the 3-year group, the scores were TNSS(0.90±0.97), VAS (1.30±1.19), CSMS (1.70±1.28); the scores of patients who completed the immunotherapy for 24 months in 2-year group and 3-year group were TNSS (2.10±0.95, 2.00±0.97), VAS (3.00±1.56, 3.10±1.68) and CSMS (3.10±1.15, 2.90±1.19) and the patients who completed 12-month immunotherapy were scored in 1-year group, 2-year group and 3-year group with TNSS(3.20±1.27, 3.10±1.41, 3.20±1.41), VAS(4.50±2.11, 4.70± 2.19, 4.50±2.17) and CSMS(4.20±1.39, 3.70±1.32, 4.10±1.39) respectively; patients with poor efficacy in sublingual immunotherapy achieved a score similar to the control group after 6 months of dose adjustment (equals to 12 months after treatment), that were CSMS(2.90±1.56, 2.90±1.88, 2.40±1.69), TNSS(4.70±2.98, 3.90±2.77, 3.80±2.45) and VAS(4.20±1.29, 4.50±1.65, 4.20±1.14) of 4 drops group, 5 drops group and control group; sublingual immunotherapy for patients with multiple allergens for 2 years finally achieved similar efficacy to patients with single allergen, with CSMS (2.30±0.50, 2.10±1.01, 1.90±1.01), TNSS (3.50±2.62, 3.70±2.62, 3.20±2.82) and VAS (4.50±1.00, 4.10±1.57, 3.80±1.54) in single allergen group, combined with 1-2 allergens group and combined with 3+ allergens group.Conclusions:Sublingual immunotherapy plays a corresponding role in the treatment of low-age children, multiple allergy children, and some children get better after dose adjustment.

OBJECTIVETo explore the relationship between NB4 cell apoptosis induced by tanshinone IIA (TanIIA) and the cell mitochondrial transmembrane potential (DeltaPsim).

METHODSNB4 cells were treated with TanIIA, As(2)O(3), TanIIA plus 1.0 micro g/ml CsA and As(2)O(3) plus 1.0 micro g/ml CsA, respectively. Morphological changes were observed under light microscope and transmission electron microscope. The percentages of sub-G(1) cells and DeltaPsim of cells doublely stained with PI and Rh123 were assayed by flow cytometry.

RESULTSThe percentages of sub-G(1) cells after treatment with 1.0 micro g/ml and 2.0 micro g/ml TanIIA had no significant difference but was higher than that of 0.5 micro g/ml. After treatment with TanIIA, NB4 cells appeared the classical apoptotic morphology. The percentages of sub-G(1) cells were increased, while the DeltaPsim reduced (P < 0.01) and there was a linear correlation between them. The increment of sub-G(1) cell percentages and decrement of DeltaPsim induced by TanIIA were partly inhibited by CsA (P < 0.01).

CONCLUSIONSTanIIA can induce NB4 cells apoptosis through opening the mitochondrial permeability transition pore and reducing DeltaPSgr;m, and this effect could be inhibited by CsA.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; DNA ; metabolism ; Diterpenes, Abietane ; Humans ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; Phenanthrenes ; pharmacology ; Tumor Cells, Cultured

This study examined the effects of flow shear stress on the bio-capacity of the endothelial cells' induced from mesenchymal stem cells (MSCs). After cultivating the SD rat mesenchymal stem cells in vitro, we exposed them under different intensity of flow shear stress and induced these cells to endothelial cells. The variations of total anti-oxidation competence (T-AOC) and quantity of nitrogen monoxide (NO) were tested. The results showed that shear stress has an enhanced effect on the T-AOC and NO of endothelial cells induced from MSCs in an intensity-dependent manner. Flow shear stress could provide a protective action on the in vitro induction of endothelial cells, thus formulating a theoretical foundation for the therapeutics of ischemic heart diseases and vascular tissue engineering.
Animals ; Antioxidants ; metabolism ; Bone Marrow Cells ; cytology ; physiology ; Cell Differentiation ; physiology ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; physiology ; Oxidation-Reduction ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical

Purpose: This study aimed to elucidate whether lncRNA ZFAS1 is involved in neuronal apoptosis and inflammation in temporal lobe epilepsy (TLE). Materials and Methods: Ninety-six TLE patients were recruited, and their peripheral venous blood was gathered to determine Zfas1 expression with polymerase chain reaction. Neurons were separated from hippocampal tissue of newborn SD rats, and siZfas1 or pcDNA3.1-Zfas1 was transfected into the neurons. Inflammatory cytokines released by neurons were determined, and neuronal activities were evaluated through MTT assay, colony formation assay, and flow cytometry. Results: Serum levels of Zfas1 were higher in TLE patients than in healthy controls (p<0.05). Furthermore, Zfas1 expression in neurons was raised by pcDNA3.1-Zfas1 and declined after silencing of Zfas1 (p<0.05). Transfection of pcDNA-Zfas1 weakened the viability and proliferation of neurons and increased neuronal apoptosis (p<0.05). Meanwhile, pcDNA3.1-Zfas1 transfection promoted lipopolysaccharide-induced release of cytokines, including tumor necrosis factor-α, interleukin (IL)-1, IL-6, and intercellular adhesion molecule-1 (p<0.05), and boosted NF-κB activation by elevating the expression of NF-κB p65, pIκBα, and IKKβ in neurons (p<0.05). Conclusion Our results indicated that lncRNA ZFAS1 exacerbates epilepsy development by promoting neuronal apoptosis and inflammation, implying ZFAS1 as a promising treatment target for epilepsy.