Objective To investigate the clinical effect of milrinone in the treatment of children with congenital heart disease complicated by severe pneumonia and heart failure. Methods 94 patients with congenital heart disease complicated with severe pneumonia and heart failure in our hospital from March 2014 to February 2016 were selected,and randomly divided into experimental group and routine group,47 cases in each groups. The routine group were treated with conventional treatment, the experimental group in the conventional group received routine treatment based on the same given milrinone treatment. The clinical efficacy, symptom disappearance time and adverse reactions were compared between the two groups. Results The treatment in the experimental group the total efficiency (93.62%) was significantly higher than that of the conventional treatment group with the total effective rate (78.72%), significant difference, statistically significant (P<0.05); the experimental group in children with fever, cough, rales and other symptoms disappeared time is lower than the conventional group, the data had significant difference, has statistical significance (P<0.05); two groups of patients during treatment were not any serious adverse reactions, the adverse reaction condition no difference does not have statistical significance.Conclusion The clinical effect of milrinone in the treatment of children with congenital heart disease complicated by severe pneumonia and heart failure significantly, clinical efficacy, low incidence of adverse reactions, worthy of promotion.

OBJECTIVE: To provide a reference about choosing the methods of isolating exosomes derived from tumor cells including laryngocarcinima Hep-2 cells by comparing advantages and defects of two methods of isolation and extraction exosomes. METHOD: Previously, laryngocarcinoma Hep-2 cells were cultivated massively, then the cells were processed with hot shock in 42 degrees C for 1 h. Sucrose density gradient centrifugation ultrafiltration (method 1): cells culture supernatant 90 ml was gathered, the supernatant was clarified through a 3/0.8 μm small filter to remove impurities and fragments which in larger diameter. Then the filtering fluid was concentrated and purified through sucrose density gradient centrifugation and ultrafiltration, the concentrated fluid was obtained. Exosome Isolation Kit (method 2): cells culture supernatant 4 ml was gathered, the solutions of the kit were added into the supernatant in proper sequence, then filtered by the special column, the concentrated fluid was obtained. Both products are observed by high resolution transmission electron microscopy. RESULT: Both methods could isolate and extract exosomes feasibly. In single high power view of transmission electron microscopy, exosomes of method 1 disperse better, but lower density, and more impurity in background, exosomes of method 2 arrange closer, higher density, and less impurity. CONCLUSION Exosome isolation Kit require less supernatant, cost less time, process procedure briefly, harvest higher yield. It may become a new option of isolating exosomes derived from Laryngocarcinoma Hep-2 cells.
Cell Line, Tumor ; Exosomes ; ultrastructure ; Humans ; Laryngeal Neoplasms ; pathology ; Microscopy, Electron, Transmission

[ABSTRACT]OBJECTIVETo investigate the expressions of Endoglin (CD105) and Galectin-3 protein in laryngeal squamous cell carcinoma (LSCC) as well as the relationship between their expressions and the clinicopathological factors of LSCC.METHODSThe expressions of CD105 and Galectin-3 protein were detected in 76 samples of LSCC and 25 normal laryngeal tissues (NLT) by immunohistochemical staining (S-P).RESULTS 1.The mean of Microvessel density (MVD) value marked by CD105 in LSCC was 10.33±2.29, which was significantly higher than that in NLT (1.20±1.04) (P<0.05). The expression of MVD marked by CD105 (CD105-MVD) was correlated with histological grading, T stage, clinical stage, lymph node metastasis, recurrence and prognosis in LSCC (P<0.05). 2.The positive expression rate of Galectin-3 protein in LSCC was 86.84%, which was significantly higher than that in NLT (36%)(P<0.05). The expression of Galectin-3 was correlated with T stage,clinical stage, lymph node metastasis, recurrence and prognosis in LSCC (P<0.05). 3.There was a positive correlation between CD105 and Galectin-3 protein. 4.Survival analysis indicated that the expressions of CD105 and Galectin-3, histological grading, lymph node metastasis,T stage and recurrence were independent factors for tumor prognosis in LSCC (P<0.05). CONCLUSIONThe expressions of CD105 and Galectin-3 protein have a positive correlation in LSCC. They may play important roles in the tumorigenesis, malignant progression and poor prognosis of LSCC. Combined detection of them may be great value in diagnosis and predicting prognosis of LSCC.

Objective To investigate the clinical effect of Zero-profile anterior cervical interbody fusion on single-segment cervical spondylosis in the elderly, and to compare with the effect of traditional anterior cervical decompression and fusion (ACDF) with titanium plate.Methods Clinical data of elderly patients who underwent anterior cervical discectomy and fusion operation (one-or two-level) in our department from June 2009 to March 2014 were retrospectively analyzed.The 49 patients who took anterior cervical interbody fusion with the Zero-profile cage were considered as the Zero-P group, and the other 60 patients who received ACDF with titanium plate as control (ACDFP group).The operation time, blood loss, Neck Disability Index (NDI) score before and after operation, Cobb angle of the cervical spine and Cobb angle of the operated segment before and after operation, the fusion rate 1 year after operation, the dysphagia rate after operation according to Bazaz index were recorded and assessed.Results The operation time and blood loss had no statistical difference between the two group (P＞0.05 for both).The NDI scores were declined in the two group after operation as compared with before treatment (P＜0.05 for both), while there were no statistical difference in NDI score between the two groups (P＞0.05).The Cobb angle of cervical spine had little improvement 1 day after operation as compared with before operation, and improved significantly 3 month after operation in the two group.The Cobb angle of operated segment (Cobb S) was reduced 3 months in the two group after operation as compared with before treatment, and the Cobb S was declined much more in Zero-P group than in control group (P＜0.01).There was no statistical difference in the number of patients with successful fusion between the two group (46 cases vs.56 cases, x2 =0.08, P＞ 0.05).Conclusions The clinical effect of Zero-profile anterior cervical interbody fusion is equivalent as the traditional ACDFP.Zero-profile anterior cervical interbody fusion has less exposure and blood loss, which is more suitable for the elderly patients with cervical degenerative disease.

Objective To explore the surgical skills and evaluate the efficacy of single small incision herniorrhaphy for bilateral inguinal hernia. Methods From August 2006 to January 2013, 26 patients with bilateral inguinal hernia received single small incision herniorrha-phy,and the clinical data,including the operation time, hospital stay,postoperative complications and the recurrence,were analyzed retrospec-tively. Results In 26 cases of bilateral inguinal hernia,the operation time was (50. 6 ± 11. 6) min,the postoperative hospital stay was (5. 8 ± 1. 4) d,the incidence of postoperative complications was 3. 8%. No recurrence occurred during the follow-up of 10 to 58 months. Conclu-sion The single small incision herniorrhaphy for bilateral inguinal hernia is proved to be effective,safe and convenient,and it has few post-operative complications. This method is worth popularizing in the clinical practice.

Objective To build convenient structure ,efficient and intelligent transfusion information system ,improve the quality of services to meet the needs of clinical applications and hospital management ,and make the core concept of “quality safety” practi‐cable by the information system .Methods Establish a blood transfusion information system based on the theory of hospital bionic management in hospital .Results The transfusion information system which constructed from bionic management theory can fully realize the design goals and satisfy all types of users .Conclusion Bionic management with the features of integrity ,auto correla‐tion ,ideology and culture has good potential for growth ,and could play a great directive role on hospital information system .

Objective:To construct tumor cell model by determination of the pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid expressing steadily in mouse melanoma B16 cells.Methods:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid being constructed from the melanoma-associated antigen A3 genes sourcing laryngocarcinoma in advance was translated into the mouse melanoma B16 cells under the mediation of lipofectamine,and the positive clones were detected with G418.The expression of enhanced green fluorescent protein( EGFP) and MAGE-A3 mRNA in positive clones were detected by fluorescence microscopy and fluorescence quantitative PCR ( qRT-PCR ) assay, respectively.Results:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected into B16 cells successfully, the green fluorescence of fusion protein expression was found, and MAGE-A3 mRNA transcription in B16 cells expressions were detected in positive clones.Conclusion:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected effectively and expressed stably by liposome method in the B16 cells.The expression of MAGE-A3 tumor cell model has been successfully established,which provide data for the study of laryngocarcinoma immunotherapy.

Objective To understand the changes of allergic rhinitis from immunology,pathophysiology and morphology method were applied.Methods Toluene 2,4-diisocyanate was dissolved with florence oil to final concentration 10%,this solution was dropped into the nasal vestibules of animals to induce sensitization process.8 guinea pigs were prepared as allergic rhinitis with 10% Toluene 2,4-diisocyanate,and other 8 animals were used as blank controh.After model successfully established,the blood were obtained to determine RBC-C3b receptor rosette rate and RBC-imuuunocomplex rosette rate.The nasal mucosas were obtained from 2 groups,distribution and changes of substance P in nasal mucosa were observed with the application of immunohistochemical staining.The content of blood histamine was determined with ELASA method.The pathological changes of nasal mucosas were observed with the application of light microscope and transmission electron microscope.Results The behavior scores of the modeling animals were significantly higher than that of controls(P＜0.01).The value of RBC-C3b reeeptor rosette rate and RBC-imuuunoeomplex rosette rate of the modeling animals were significantly decreased than that of the controls(P＜0.05).Substance P presented in the normal nasal mucosal epithelial cells,epithelium cells of blood vessels,glandular cells and its duets,the staining density and the positive staining cells in the same aero in modeling group significantly increased,compared with controls.The counts of substance P-positive cells of control group were less than those of modeling group(P＜0.05).The content of blood histamine of the modeling animals were significantly increased compared with the controls(P＜0.01).The structure of false multiple coat cilium columnar epithelial cells in nasal mucosa of control group were successive,intact,and distinct.There were normal mucosal epithelium,lamina propria and submucosa.But modeling group showed that mucosal epithalamiums were damaged and shed,goblet cell proliferated,squamous metaphase,and epithelial necrosis happened,serous glands in lamina propria vigorously proliferated,blood vessels expanded,tissue edema formed,plenty of inflammatory cell such as eosinophil and mast cells were more in number and infiltrated.The structure of epithelial eells,cilia and mierotubule of nasal mucosa of control group were regular and distinct,there were abundant cellular organs in cytoplasm.Eosinophil cells were intact.But iu model group,mucosal epithalamium,cilia and its microtubule,mierovillus,goblet cell were damaged,the cell bulk and nucleus changed,mast cells and eosinophil cells changed,blood vessels expanded,serous glands vigorously proliferated.This morphological change was roughly identical to clinical manifestation of allergic rhinitis.Conclusion Toluene-2,4-diisocyanate can be used to establish allergic rhinitis model in guinea pigs,and some ehanges of the allergic rhinitis in model guinea pig were similar with clinic observation.

OBJECTIVE: To study the assistant effect of Scutellaria barbata extract (ESB) in 5-fluorouracil (5-FU) chemotherapy. METHODS: A mouse model of transplanted hepatoma H22 was used in this study to evaluate the synergic and attenuating effects of ESB in chemotherapy. Tumor inhibition rate, life span of mice and the toxicity of chemotherapy were observed. The body weight, tumor weight, thymus index and spleen index in H22-bearing mice were also measured. The phagocytotic function of macrophages was studied by observing phagocytization of peritoneal macrophages. RESULTS: The increase of body weight in 5-FU plus ESB groups was higher than that in 5-FU group, and the side effects such as anorexia, abdominal distention and athrepsy were relieved. Compared with untreated group, prolonged lifetime in 5-FU plus high-dose ESB group and 5-FU plus low-dose ESB group was improved. Life prolongation rates in 5-FU plus high-dose ESB group and 5-FU plus low-dose ESB group were 61.46% and 23.59% respectively. High-dose ESB, 5-FU, 5-FU plus low-dose ESB and 5-FU plus high-dose ESB could inhibit the tumor growth, and the tumor inhibition rates were 36.98%, 42.26%, 52.45% and 65.28%, respectively. Thymus index and spleen index were increased significantly in 5-FU plus low-dose ESB group and 5-FU plus high-dose ESB group. White blood cell (WBC) count was decreased obviously in 5-FU group, while the count of WBC was increased in 5-FU plus low-dose ESB group and 5-FU plus high-dose ESB group. The phagocytotic function of macrophages was also increased in 5-FU plus low-dose ESB group and 5-FU plus high-dose ESB group. CONCLUSION: ESB can enhance the effect of chemotherapy, relieve the side effects and improve immune function of mice in chemotherapy. These results suggest that ESB, as a biochemical modulator to enhance the therapeutic effects, is useful in cancer chemotherapy.

Objective To investigate the methods and results of perforator pedicled flaps by anastomosis of superficial veins for reconstructing the extremity defects.Methods From February, 2012 to September, 2012, a total of 14 patients with traumatic skin and soft tissue and artery defects in extremities were repaired by perforator pedicled flaps anastomosed superficial veins with the recipient vessels.Fourteen flaps were based on five kinds of perforator arteries, the posterior interrosseous artery in 5 cases, the ulnar artery in 3 cases, the metacarpal artery in 2 cases, the peroneal artery in 3 cases, the lateral supramalleolar artery in 2 cases, the ulnar artery in 2 cases,and the posterior interrosseous artery in 1 case.After the operation, the blood supplies of flaps were observed severely,postoperative flap swelling were evaluated by edema level classification.After 1 month the blood flow of vein anastomosis were examed by color Doppler ultrasound.Questionnaire of the flap aesthetic satisfactory were performed during fellow-up periods.Results Twelve flaps were all survived well, 2 flaps had partial marginal skin necrosis in the distal, which was managed with surgical debridement, and wound healed in 1 month.Flap swelling were light, edema level classification were one degree in 4 cases,two degree in 10 cases postoperation 7 days, one degree in 11 cases and two degree in 3 cases, after 6 months.Color Doppler ultrasound examinations showed 9 vein anastomosises were all bypassed well.Eleven cases had 6-12 months' fellow-up periods.The flaps had like-like appearance, good contour, high aesthetic satisfactory.Conclusion Perforator pedicledr flaps by anastomosis of superficial veins can reduce the flap venous pressure obviously, avoid transientvenous venous congestion, improve the survival quality of the flap.It's a safe and effective method for soft tissue coverage of traumatic extremity wounds.