ObjectiveTo analyze the effect of temporal distance parameters on comfortable and maximal walking speed of hemiplegic stroke patients.MethodsThe comfortable and maximal walking speed of 85 hemiplegic stroke patients were tested by 10 m walking speed and temporal distance parameters of gait cycle were obtained. The effect of step length and walking rate on comfortable and maximal walking speed was analyzed.ResultsStep length and walking rate were significantly positive related to comfortable and maximal walking speed (r=0.849-0.915,P<0.001).The step regression analysis selected step length as a significant variable for comfortable and maximal walking speed (R2=0.835,R2=0.827,respectively). ConclusionThe important parameter that influences comfortable and maximal walking speed of hemiplegic stroke patients is step length.

A total of 121 subjects comprising 40 normal subjects,58 patients with overweight or obesity and 23 patients with Cushing's syndrome were recruited in the study.The modified radioimmunoassay (RIA) for salivary cortisol test was established'and its normal range was determined.Then the diagnostic value of the salivary cortisol for the initial diagnosis of Cushing's syndrome was evaluated and single midnight salivary cortisol test demonstrated a sensitivity of 100.0% and specificity of 91.4 %.Salivary cortisol test can be recommended as a first-line diagnostic parameter for Cushing's syndrome.

OBJECTIVES: Chronic rhinosinusitis (CRS) is common disease in otorhinolaryngology and will lead to lower airway abnormality. However, the only lung function in CRS patients and associated factors have not been much studied. METHODS: One hundred patients with CRS with nasal polyps (CRSwNP group), 40 patients with CRS without nasal polyps (CRSsNP group), and 100 patients without CRS were enrolled. The difference in lung function was compared. Meanwhile, CRSwNP and CRSsNP group were required to undergo a bronchial provocation or dilation test. Additionally, subjective and objective outcomes were measured by the visual analogue scale (VAS), 20-item Sino-Nasal Outcome Test (SNOT-20), Lund-Mackay score, Lund-Kennedy endoscopic score. The correlation and regression methods were used to analyze the relationship between their lung function and the above parameters. RESULTS: The forced expiratory volume in 1 second (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25-75) of CRSwNP group were significantly lower than other groups (P<0.05). On peak expiratory flow, there was no difference between three groups. In CRSwNP group, FEV1 was negatively correlated with peripheral blood eosinophil count (PBEC) and duration of disease (r=–0.348, P=0.013 and r=–0.344, P=0.014, respectively), FEF25-75 negatively with VAS, SNOT-20 (r=–0.490, P=0.028 and r=–0.478, P=0.033, respectively) in CRSsNP group. The incidence of positive bronchial provocation and dilation test was lower in CRSwNP group (10% and 0%, respectively), with both 0% in CRSsNP group. The multiple linear regression analysis indicated that change ratio of FEV1 before and after bronchial provocation or dilation test were correlated with PBEC in CRSwNP group (β=0.403, P=0.006). CONCLUSION: CRS leading to impaired maximum ventilation and small airway is associated with the existence of nasal polyp. Lung function impairments can be reflected by PBEC, duration, VAS, and SNOT-20. In CRSwNP patients, PBEC is independent predictor of FEV₁ change ratio.
Bronchial Hyperreactivity ; Bronchial Provocation Tests ; Eosinophils ; Forced Expiratory Volume ; Humans ; Incidence ; Linear Models ; Lung* ; Nasal Polyps ; Otolaryngology ; Ventilation ; Vital Capacity

OBJECTIVETo explore the effect of Toll-like receptor 4 (TLR4) activation on the migration of asthmatic airway smooth muscle cell (ASMCs) induced by airway epithelial cells.

METHODSPrimary ASMCs were cultured by the method of cell digestion. Cell culture supernatant of RTE cells were collected by TNF-alpha stimulation of epithelial cells. Detected the IL-8 and RANTES levels in the supernatant. The transmembrane migration of asthmatic ASMCs were detected by Modified Boyden chemotaxis chamber. The effect of TLR4 on the migration of asthmatic ASMCs induced by epithelial cells with TLR4 antibody drugs as a tool.

RESULTSThe levels of IL-8 and RANTES in the supernatant of TNF-alpha groups were significantly increased, and that in the 20 ng/ml group was significantly higher than other groups (P < 0.01). The transmembrane migration of asthmatic ASMCs groups was increased than that of control group. The transmembrane migration of asthmatic ASMCs from asthma group and TNF-alpha + TLR4 antibody group was significantly decreased compared with that in TNF-alpha group (P < 0.01). The migration of asthma ASMCs from TNF-alpha + TLR4 antibody group was increased than that of asthma group (P < 0.05).

CONCLUSIONTLR4 in the surface of asthmatic ASMCs may be activated by cytokines secreted by the airway epithelial cells and enhance the transmembrane migration of asthmatic ASMCs induced by airway epithelial cells so that it plays a role in airway remodeling of asthma.

Animals ; Asthma ; metabolism ; Cell Movement ; Cells, Cultured ; Chemokine CCL5 ; metabolism ; Epithelial Cells ; metabolism ; Interleukin-8 ; metabolism ; Myocytes, Smooth Muscle ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVEThe abnormality of hemopoietic inductive microenvironment (HIM) is involved in the pathophysiology of aplastic anemia (AA). Mesenchymal stem cells (MSC) are main source of bone marrow stromal cells which constitute the bone marrow HIM. Thus, the bone marrow failure in AA may be related to the function of MSC. The aim of the study was to investigate the hematopoiesis support function of MSC in children with AA in vitro.

METHODSBone marrow samples were collected from 24 children with AA at diagnosis and 19 children with idiopathic thrombocytopenic purpura (ITP), infectious mononucleosis or lymphadenitis (controls). MSCs from bone marrow samples were isolated, cultured and expanded. Morphology, proliferation activity and colony forming unit-fibroblast (CFU-F) were measured. The ability of bone marrow MSC to adhere hemopoietic cells was assayed by MTT. The concentration of stem cell factor (SCF) released from MSC was tested using ELISA. Mononuclear cells (MNC) of bone marrow were plated onto a feeder layer formed by MSC. Cells count and BFU-E, CFU-GM, CFU-GMME productions were measured.

RESULTSThe first and third passage time of MSC in children with AA was longer than that in the controls. The number of CFU-F in children with AA (15.70+/-5.78) was less than that in the controls (21.73+/-5.74) (P<0.05). The concentration of SCF in MSC supernatants in children with AA (30.69+/-16.82 pg/mL) was significantly lower than the controls (50.74+/-14.83 pg/mL) (P<0.01). The total MNC count and the number of BFU-E, CFU-GM and CFU-GMME colonies in the support of MSC in children with AA were significantly lower than those in the controls (P<0.01).

CONCLUSIONSThe hematopoiesis support function of MSC was significantly reduced in children with AA in vitro. The decreased hematopoiesis support function of MSC may be related its decreased proliferation capacity and SCF release activity.

Adolescent ; Anemia, Aplastic ; physiopathology ; Cell Adhesion ; Child ; Child, Preschool ; Female ; Hematopoiesis ; Humans ; Leukocytes, Mononuclear ; physiology ; Male ; Mesenchymal Stromal Cells ; physiology ; Stem Cell Factor ; physiology

OBJECTIVETo study the influence of recombinant lentiviral vector encoding miR-15a/16-1 on biological features of human nasopharyngeal carcinoma CNE-2Z cells and underlying mechanisms.

METHODSGFP-positive CNE-2Z cells transfected with recombinant lentiviral vector were selected. The experiment was divided into control group, transfected group, radiotherapy group, transfected-radiotherapy group. Cell proliferation was analyzed by MTT. Apoptosis was detected by flow cytometry. Radiotherapy sensitivity of the cells in control group and transfected group was evaluated by colony forming experiment. The expressions of miR-15a, miR-16-1 and bcl-2 mRNA were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The expression of bcl-2 protein was detected by Western blot. The activation of Caspase-2 and Caspase-3 was evaluated by spectrophotometry.

RESULTSRelative expression quantities of miR-15a and miR-16-1 in infected group were 524.80 ± 40.79 (t = 494.611, P = 0.000) and 466.11 ± 40.96 (t = 386.8, P = 0.000), respectively. The proliferation of the cells in transfected-radiotherapy group was the most obvious, followed by the cells in radiotherapy group and transfected group (F = 424.3, P = 0.000). The apoptosis rates of control group, transfected group, radiotherapy group and transfected-radiotherapy group were (2.2 ± 1.4)%, (9.6 ± 0.8)%, (2.9 ± 1.1)%, and (18.6 ± 0.7)% respectively(F = 158.5, P = 0.000). Clonogenic assay showed that the values of SF2, Do (1.473) and Dq (1.581) in transfected group were lower than those in control group. The relative expression levels of bcl-2 mRNA in transfected group, radiotherapy group, and transfected-radiotherapy group had no significant difference (P > 0.05). Decrease in the expression of bcl-2 protein in transfected-radiotherapy group was most significantly, followed by that in transfected group. The percentages of activated Caspase-2 in control group, radiotherapy group, transfected group and transfected -radiotherapy group were 0.12 ± 0.01, 0.24 ± 0.04, 0.35 ± 0.02, and 0.44 ± 0.04, respectively (F = 115.500, P = 0.000). The percentages of activated Caspase-3 in the groups were 0.13 ± 0.01, 0.27 ± 0.01, 0.43 ± 0.02, and 0.83 ± 0.06, respectively (F = 439.921, P = 0.000).

CONCLUSIONSRecombinant lentiviral vector LV-miR15a/16-1 could improve the expression of miR-15a and miR-16-1 in CNE-2Z cells, inhibit the proliferation of CNE-2Z cells, promote apoptosis and enhance the sensitivity of the cells to radiotherapy probably by inhibiting bcl-2 expression, activating Caspase-2 and Caspase-3.

Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Carbamates ; Carcinoma ; Caspase 2 ; metabolism ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cysteine Endopeptidases ; metabolism ; Genetic Vectors ; Humans ; MicroRNAs ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; Pyrazoles ; RNA, Messenger ; Strobilurins ; Transfection

OBJECTIVETo study the effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering.

METHODSBased on the colon preference of E. coli, the HDV small antigen original gene from GenBank was optimized. Both the original gene and the optimized gene expressed in prokaryotic cells, SDS-PAGE was made to analyze the protein expression yield and to decide which protein expression style was more proportion than the other. Furthermore, two antigens were purified by chromatography in order to compare the purity by SDS-PAGE and Image Lab software.

RESULTSSDS-PAGE indicated that the molecular weight of target proteins from two groups were the same as we expected. Gene optimization resulted in the higher yield and it could make the product more soluble. After chromatography, the purity of target protein from optimized gene was up to 96.3%, obviously purer than that from original gene.

CONCLUSIONGene optimization could increase the protein expression yield and solubility of genetic engineering HDV small antigen. In addition, the product from the optimized gene group was easier to be purified for diagnosis usage.

Electrophoresis, Polyacrylamide Gel ; Genetic Engineering ; methods ; Hepatitis D ; diagnosis ; Hepatitis delta Antigens ; genetics ; isolation & purification ; Recombinant Proteins ; biosynthesis ; isolation & purification

OBJECTIVETo identify differentially expressed genes in recurrent nasopharyngeal carcinoma (rNPC) by DNA microarrays, and analyze chromosomal localizations and molecular function by bioinformatics.

METHODSThe primary nasopharyngeal carcinoma (pNPC) tissue samples and rNPC tissue samples were selected, and Affymetrix Gene1.0 ST gene chips were used to identify differential expressed genes in rNPC, and the bioinformatics was used to analyze their chromosomal localizations as well as molecular functions.

RESULTSA total of 44 genes were identified to be differential expressed in rNPC. Thirty-six genes were down regulated, 8 genes were up regulated. Functional classification of down-regulation genes showed that most genes (10 genes, 27.8%) belonged to the enzyme activity genes, followed by calcium ion binding genes (7 genes, 19.4%), protein binding genes (5 genes, 13.9%), receptor activity genes (4 genes, 11.1%), ATP binding genes (2 genes, 5.6%), transcription factor genes (2 genes, 5.6%), extracellular matrix binding and growth factor binding have 1 gene respectively (each accounted for 2.8%). In addition, the functions of 4 genes (11.1%) were unknown. Functional classification of up-regulation genes showed most genes (3 genes, 37.5%) were unknown, followed enzyme activity genes (2 genes, 25.0%), receptor activity, calcium ion binding and voltage-gated ion channel activity genes have 1 genes respectively (each accounted for 12.5%). These genes were localized randomly on the most the chromosomes, with a majority of them localized on chromosomes 1, 17. Chromosome 1 contained the most differentially expressed genes (10, 22.7%), followed by chromosomes 17 (5, 11.3%).

CONCLUSIONSThe differential expressed genes in rNPC were supposed to be randomly distributed on most chromosomes, but the majorities were found on chromosomes 1, 17. Abnormality in three groups of genes, including in enzyme activity, calcium ion binding and protein binding associate genes, might play important roles in rNPC. Those genes need to be further studied.

Adult ; Aged ; Carcinoma ; Carcinoma, Squamous Cell ; genetics ; pathology ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Neoplasm Recurrence, Local ; genetics ; Oligonucleotide Array Sequence Analysis

Objective To study the effect of mesenteric lymph duct ligation(MLDL)on sepsis-induced lung and intestinal injuries in rats.Methods Sepsis was induced by cecal ligation and puncture(CLP)in male rats.The rats were randomly divided into sham control group in which rats received sham operation,sepsis group(CLP group)in which rats received saline after CLP operation,and CLP+MLDL group in which rats received mesenteric lymph duct ligation(MLDL)after CLP operation.The rats were sacrificed at 24 h after CLP modeling. Bronchoalveolar lavage fluid,arterial blood,and intestine and lung tissues were collected to determine organ injuries.Results The lung tissue histopathology and blood gas analysis revealed that MLDL markedly alleviated the lung injury(ALI score:6.14 ± 0.51 vs.8.12 ± 0.63,P=0.029 9),improved oxygen partial pressure[(78.67 ± 4.51)mmHg vs.(64.83 ± 2.90)mmHg,P=0.0273],decreased lung W/D ratio(6.12 ± 0.25 vs.7.63 ± 0.49,P=0.021 2),decreased BALF cytokines levels[TNF-α:(828.17 ± 81.89)pg/mL vs.(1 118.17 ± 79.22)pg/mL,P=0.029 1;IL-6:(39.33 ± 5.50)ng/mL vs.(68.67 ± 5.24)ng/mL,P=0.003 4],alveolar permeability[protein levels:2.117 ± 0.289 2 vs.3.033 ± 0.164 7,P=0.020 3,and cell levels:(30.00 ± 3.587)×106/L vs.(43.83 ± 2.358)×106/L,P=0.009 1].However,according to the results of HE and biochemical detection,MLDL had no protective effect on sepsis-induced intestinal injury.Moreover,MLDL could significantly reduce the bacterial loads of the blood and lung,but do not change the bacterial level in the intestines.Conclusion MLDL has a significant protective effect on sepsis-induced lung injury mainly by decreasing bacterial translocation,but had no effect on intestinal injury.This difference may be related to that MLDL inhibits the bacteria spreading from abdominal cavity to other organs in sepsis but it causes their accumulation in the intestines.

Objective To discuss the optimal clinical diagnosis and treatment of ectopic ACTH syndrome with occult tumors. Methods Clinical features, imaging examinations and treatment of 17 patients with ectopic ACTH syndrome were described and compared. Results All patients illustrated the typical clinical features of Cushing’s syndrome. They had hypokalemic alkalosis, elevated serum cortisol and plasma ACTH levels. In the high-dose dexamethasone suppression tests, most patients failed to suppress serum cortisol and 24-hour urinary cortisol. CT and MRI are useful imaging modalities to localize the ACTH-secreting tumor in patients with ectopic ACTH syndrome. The patients with overt ACTH-secreting tumors had surgical curative resection soon after diagnosis. Among patients with occult ACTH-secreting tumors, three underwent subtotal bilateral adrenalectomy, two underwent right adrenalectomy, four received inhibitor of steroidogenesis aminoglutethimide. Their hypercortisolism was controlled. Conclusion Surgical curative resection is the optimal treatment of ectopic ACTH syndrome with overt ACTH-secreting tumor. Bilateral adrenalectomy, right adrenal ectomy or chemotherapy to control hypercortisolism is an available treatment of ectopic ACTH syndrome with occult ACTH-secreting tumors.