Objective To investigate the phenotypes and expression of activated T-cell membrane molecules on lymphocytes in the peripheral blood(PB) of children with aplastic anemia (AA) and find out its clinical significance.Methods The lymphocytes from PB of the 115 AA patients were analyzed by flow cytometry and compared with normal group.Results The expression of CD8+ increased and the expression of CD4+ decreased. The ratio of CD4/CD8 cells decreased ,and was more significant in the patients with serious AA(SAA). The expression of HLA-DR and ??T-cell also increased, and was more significant in SAA patients. The expression of CD3+ was not significantly changed.Conclusions There is significant immune abnormal in AA patients and more important in SAA patients. The immunosuppressant is useful in clinical treatment.

Zebrafish (Danio rerio) is a simple biological labora-tory animal, with similar biological structure and physiological functions to mammals. Almost transparent embryos, in vitro de-velopment of embryos and up to 87% of human genetic similari- ties enable a wide application of zebrafish in the field of pharma-ceutical research. In this paper, we have studied the application of zebrafish in drug metabolism, the research of Chinese medi-cine, the evaluation of drug toxicology and safety, the screening of drugs and the discovery of new drugs, the research of regener- ative drugs, so as to provide new ideas of zebrafish in the field of pharmacy.

OBJECTIVETo investigate the difference of soluble P-selectin levels in different subtype of coronary heart disease and the relationship between soluble P-selectin levels with the severity of coronary artery lesions.

METHODSEnzyme linked immuoserbent assay (ELISA) was used to measure the plasma soluble P-selectin levels in 69 patients with angiocardiography documented coronary heart disease and 19 normal coronary arteries persons without angiocardiography detectable coronary artery disease (control group). The coronary artery lesions score was recorded according to single, double and triple-vessel lesions while the American College of Cardiology and the American Heart Association proposed type A, B, C lesion and Gensini scoring system. The relationships between plasma soluble P-selectin levels and the coronary artery score (the severity of coronary heart disease) were assessed.

RESULTS(1) The level of plasma soluble P-selectin was obviously higher in the coronary heart disease group than in the control group (180.6 +/- 60.5 ng/L vs. 145.3 +/- 21.7 ng/L, P<0.05). (2) The level of plasma soluble P-selectin was significantly higher in the acute coronary syndrome group (191.4 +/- 63.7 ng/L) than in the control group (145.3 +/- 21.7 ng/L, P< 0.01) and in the stable angina pectoris group (141.3 +/- 17.9 ng/L, P<0.01). (3) The level of plasma soluble P-selectin was high in multi-vessel coronary artery lesions group than in single-vessel group (190.1 +/- 64.2 ng/L vs. 157.2 +/- 43.4 ng/L, P < 0.05). The level of plasma soluble P-selectin was positively correlated with the Gensini score (r = 0.391, P = 0.001); the numbers of vessels lesions (rs = 0.349, P = 0.003); Type A, B and C lesions (rs = 0.358, P = 0.002).

CONCLUSIONThe positive correlation between the level of soluble P-selectin and the coronary artery score may indicate that soluble P-selectin levels might reflect the severity of coronary heart disease. The elevated soluble P-selectin level in acute coronary syndrome suggested the possible relation of P-selectin to the pathogenesis of acute coronary syndrome, which may save as a potential marker of plaque unstability.

Case-Control Studies ; Coronary Disease ; blood ; physiopathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Middle Aged ; P-Selectin ; blood ; chemistry ; Solubility

The aim of this study is to study the effect of calcium dobesilate on streptozotocin (STZ)-induced early diabetic nephrophathy (DN) in rats. All male Wistar rats were randomly divided into six groups: normal group; DN blank group; calcium dobesilate 75, 150, and 300 mg x kg(-1) groups and perindopril 0.4 mg x kg(-1) group. Blood glucose and the 24 h urinary albumin were measured dynamically during the experiment, after 8 weeks administration, the level of glycosylated hemoglobin (HbA1c) was determined, the expressions of plasminogen activator inhibitor-1 (PAI-1) and matrix metalloprotein-9 (MMP-9) in cortex of kidney were examined with immunohistochemical staining. The endothelin (ET) in plasma and kidney cortex was measured with radioimmunoassay, renal pathomorphism was observed with light and electron microscopes. Calcium dobesilate could decrease the 24 h urinary albumin and ET in plasma and kidney cortex, down-regulate the expression of PAI-1, and up-regulate MMP-9 in kidney. These findings suggested that calcium dobesilate could protect blood vessel endothelium, inhibit kidney fibrous degeneration, ameliorate renal pathological damage, and protect kidney function in many ways.
Albuminuria ; Animals ; Blood Glucose ; metabolism ; Calcium Dobesilate ; pharmacology ; Diabetes Mellitus, Experimental ; blood ; metabolism ; pathology ; Diabetic Nephropathies ; blood ; metabolism ; pathology ; Endothelins ; blood ; metabolism ; Glycated Hemoglobin A ; metabolism ; Hemostatics ; pharmacology ; Kidney Cortex ; metabolism ; ultrastructure ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo observe the correlation between brain-derived neurotrophic factor (BDNF) expression in the nerve system of diabetes mellitus (DM) rats and diabetic erectile dysfunction (ED).

METHODSDM rats were induced by injecting streptozotocin and erectile function test was done by injecting apomorphine (APO) at 1 month, 2 months, 3 months and 4 months. Then the brain, lumbosacral spinal cord, thoracic and lumbar sympathetic trunks, penis and prostate were taken from the diabetic and normal rats of the same age. The BDNF positive neurons and nerve fibers were shown by immunohistochemistry or fluorescence immunohistochemistry. The number and the grey density of BDNF positive cells and fibers were detected by image analysis.

RESULTSCompared with the control group, the erection frequency of the DM rats decreased at 2 months (P <0. 05) , and significantly at 3 and 4 months (P > 0.01) , and the BDNF positive neurons and nerve fibers in the cerebral cortex, lumbosacral spinal cord, thoracic and lumbar sympathetic trunks, penis and prostate of 1-month DM rats were reduced (P <0. 05). As time went on, BDNF declined progressively.

CONCLUSIONBDNF decreases in the central and peripheral nerve system in the early stage of diabetes mellitus, wich is closely correlated with diabetic ED.

Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; Diabetes Mellitus, Experimental ; physiopathology ; Erectile Dysfunction ; physiopathology ; Male ; Nerve Tissue ; metabolism ; Rats ; Rats, Sprague-Dawley

Child, Preschool ; Electroencephalography ; Epilepsy, Rolandic ; physiopathology ; Female ; Humans ; Male ; Syndrome

Objective To discuss the clinical effect of two-dimensional oral care for patients with mechanical ventilation by orotracheal intubation,and improve the quality of oral care.Methods Seventy inpatients with mechanical ventilation by orotracheal intubation from Feb 2011 to Jul 2012 in ICU were selected and randomly divided into experimental group (n ＝ 35) and control group (n ＝ 35).Experimental group was given two-dimensional oral care (oral swab just one time before intubation,and brush teeth and suction three times a day after intubation).Control group was given traditional oral care three times a day.Throat swab specimens were gathered for bacterial colony counts before intubation,and in 4 h,12 h,24 h,48 h after intubation,respectively.Bacteria quantitative culture was performed in sputum specimens and in throat swab specimens (one time per three days) after intubation.Oropharyngeal bacteria change and the incidence rate of ventilator associated pneumonia (VAP) were observed.Results Among all 70 patients,61 cases were valid,15 cases had VAP,and 8 cases died.In the experimental group,31 were valid,4 had VAP and 3 died,while in the control group,30 were valid,11 had VAP and 5 died.The difference of the incidence rate of VAP was statistically significant (x2 ＝ 4.643,P ＝ 0.040) and the difference of the incidence rate of death was not statistically significant (x2 ＝ 0.654,P ＝ 0.473).The difference of oropharyngeal bacterial colony counts had no statistical significance between two groups before intubation (t ＝-0.563,P ＝ 0.589),while the difference of oropharyngeal bacterial colony counts had statistical significance between groups in 4 h,12 h,24 h,48 h after intubation (t＝1.957,-2.520,-3.560,-2.165,respectively; P＜0.05).Conclusions Two-dimensional oral care can effectively reduce oropharyngeal bacterial colony,and decrease the incidence rate of VAP for patients with mechanical ventilation by orotracheal intubation.Thus it is a better nursing method to improve oral care quality.

BACKGROUNDAdenosine receptors (ADORs) have been reported to play a role in experimental myopia. This study aimed to determine the distribution of ADORs in human retinal pigment epithelium (RPE) cells cultured in vitro.

METHODSHuman RPE cells (cell line D407) were cultured in vitro. ADOR mRNA in RPE was detected by reverse transcription polymerase chain reaction. ADOR protein expression in RPE was confirmed by Western blotting analysis of cell lysates. Confocal fluorescence microscopy was used to study the subcellular distribution of ADORs.

RESULTSAll four subtypes of ADORs mRNA and protein were expressed in human RPE. This was confirmed by Western blotting analysis. The ADOR subtypes were differently distributed within the cells. ADORA1 was expressed in nucleus, perinucleus and cytoplasm of RPE. ADORA2A was concentrated mainly in one side of the perinucleus and cytoplasm of RPE. ADORA2B was strongly expressed in the nucleus, perinucleus and the cytoplasm, and ADORA3 was expressed weakly in the cytoplasm of RPE.

CONCLUSIONSADORs are expressed in human RPE. The different distribution at the subcellular level suggests different functions of ADOR subtypes.

Blotting, Western ; Cell Line ; Fluorescent Antibody Technique, Indirect ; Humans ; Receptors, Purinergic P1 ; genetics ; metabolism ; Retinal Pigment Epithelium ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVESpinal muscular atrophy (SMA) is an autosomal recessive disorder that results in symmetrical muscle weakness and wasting due to degeneration of the anterior horns of the spinal cord. The clinical picture of SMA is variable and childhood SMA has been classified into 3 types on the basis of the age of onset and clinical course. The survival motor neuron (SMN) gene was mapped to chromosome 5q13. The SMN1 gene has been recognized to be responsible for SMA because of homozygous deletions or intragenic mutations in SMN1 results in childhood onset of SMA. The main objective of this study was to determine the deletion frequency of SMN1 gene and to apply gene analysis in children patients with SMA.

METHODSThe SMA patients were diagnosed and clinically typed according to the international diagnostic criteria, following up cases, and gene analysis. The PCR enzyme assay was used to detect the homozygous deletion of SMN1 gene in SMA patients. A dosage assay that combined multiplexed allele-specific PCR and DHPLC was used to determine the copy numbers of the SMN1 and SMN2 and detect SMN1 heterozygous deletion.

RESULTS(1) A total of 267 patients with SMA were diagnosed from 338 suspicious cases and 143, 82, and 42 cases were typed as types I, II, and III, with the percentages of 53.6% (143/267), 30.7% (82/267) and 15.7% (42/267), respectively. (2) Results of the present study showed that 68.5% (183/267) of SMA patients had homozygous deletions of exons 7 and 8 of SMN1 gene and 12.7% (34/267) had homozygous deletions of only exon 7 of SMN1 gene. The SMN1 heterozygous deletion was confirmed in 12.4% (33/267) of SMA patients. Non-deletion SMA patients accounted for 6.4%(17/267). The homozygous deletions of only exon 8 of SMN1 gene could not be detected. (3) The rates of homozygous or heterozygous deletion in types I and II were very similar. The rate of homozygous deletion was lower in type III than that in type I or II and rate of heterozygous deletion of type III was higher than that in types I or II.

CONCLUSION(1) The frequency and pattern of deletions in the Chinese children patients with SMA are significantly different from that observed in Caucasians populations. Further gene characterization and subtle mutations within the SMN1 gene need to be studied in order to define the molecular basis of SMA in the Chinese population. (2) The gene diagnosis is a special and non invasive method as compared with other methods. A total of 80% patients can be diagnosed through the analysis of the homozygous deletion of SMN1 gene. (3) The clinical diagnosis and gene detection need to be studied in future for the SMA patients with type III.

Adolescent ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; European Continental Ancestry Group ; genetics ; Exons ; Female ; Gene Deletion ; Humans ; Infant ; Male ; Spinal Muscular Atrophies of Childhood ; diagnosis ; genetics ; Survival of Motor Neuron 1 Protein ; genetics

OBJECTIVETo search differentially expressed proteins in serum of patients with Crohn disease.

METHODSSerum protein samples from 4 patients with Crohn disease and 8 healthy adults were recruited cross-labeled with variant CyDye, and then followed by two-dimensional differential in-gel electrophoresis (2-D DIGE), image analysis, and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTSThe 2-D electrophoresis results were compared between the Crohn disease patients and the healthy adults. The spot 1058 expression in serum of Crohn disease patients increased by 1.68 folds as compared with healthy adults (P<0.05). The protein was identified as haptoglobin by mass spectrometry.

CONCLUSIONUp-regulating expression of haptoglobin in serum of Crohn disease patients may play a role in disequilibrium of immunity system.

Adult ; Blood Proteins ; metabolism ; Case-Control Studies ; Crohn Disease ; blood ; Electrophoresis, Gel, Two-Dimensional ; Haptoglobins ; metabolism ; Humans ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization