Background Researching the pathological characteristics and components of cells in internal limiting membrane of idiopathic macular hole (IMH) has an important clinical significance for the prevention of IMH.However,the study results are still disputable.Objective This study was to investigate the histopathological features of internal limiting membrane of IMH and the types of cells inside it, and explore the pathomechanism of IMH.Methods Seven specimens of internal limiting membrane were obtained during the vitrectomy with IMH patients in the Second Affiliated Hospital of Guangzhou Medical University from February 2012 to August 2013 under the informed consent of patients.The histopathological examination was performed for the structural observation and cellular identification of internal limiting membrane.The expression and location of glial fibrillary acidic protein (GFAP), CD45 and CD44 in internal limiting membrane were examined by using immunochemistry and immunofluorescence technology.Results All the seven specimens showed continuous undulating membrane with red staining.Two specimens appeared to be uniform in thickness and few cells were distributed in the specimens.The internal limiting membranes were uneven in thickness in the other specimens with retinal pigment epithelial cells,neuroglia cells,fibrocytes,macrophages and lymphocytes in them.Immunochemistry showed the positive expression of GFAP in the outer layer of the specimens.CD45 positive cells were detected in the internal limiting membranes, and CD44 was detected in the inner layer of the specimens.Conclusions Few cells exist in the internal limiting memranes of IMH.However,neuroglia cells and CD45 positive cells emerge in the internal limiting memranes of stage 3 or above IMH eyes, indicating the proliferation of cells and immuno-inflammation response exist during the IMH development.The up-regulation of CD44 expression promotes inflammatory response of internal limiting memranes.

Animals ; Conditioning (Psychology) ; drug effects ; Extinction, Psychological ; drug effects ; physiology ; Fear ; drug effects ; physiology ; Male ; Mifepristone ; pharmacology ; Rats ; Rats, Sprague-Dawley

Adrenal Hyperplasia, Congenital ; complications ; metabolism ; Body Height ; Growth Disorders ; etiology ; Humans ; Risk Factors ; Steroid 21-Hydroxylase ; metabolism

Background Recent studies indicated that human amnion mesenchymal stem cells (hAMSCs) can be induced to differentiate into multiple types of cells in vitro,but whether the hAMSCs can differentiate into ocular surface cells has not been reported yet.Objective This study was to investigate the feasibility of inducing differentiation of hAMSCs into ocular surface cells by co-culturing with human bulbar conjunctiva fibroblasts (hBCFs).Methods This study was approved by Ethic Committee of Affiliated Second Hospital of Guangzhou Medical University.HAMSCs were isolated from placenta under the informed consent of healthy delivery women.hAMSCs were cultured,passaged and identified by detecting the expressions of CD44,CD45,CD73,CD90 in the cells with flow cytometer,osteogenesis and adipogenic differentiation experiments.Human conjunctival tissue was obtained during the eye operation under the informed consent of patients and hBCFs were isolated and cultured with explant culture.The cells were divided into the hAMSCs culture group and the hAMSCs and hBCFs co-culture group and cultivated in Transwell chambers for 7 days.The expressions of cytokeratin 19 (CK19) and α-smooth muscle actin (α-SMA) in the cells were assayed by immnofluorescence technique.Results Cultured hAMSCs showed the slender shape and cell body enlarged with passage.CD44,CD73 and CD90 were expressed in the cells,and the expression of CD45 was absent.After 3-4 weeks of osteogenesis and adipogenic induce,the cells showed red staining for alizarin and oil red O.In the co-culture group of hAMSCs and hBCFs,hAMSCs presented the epithelioid cell-like in shape and showed the positive response for CK19 and weaker response for α-SMA.However,in the hAMSCs culture group,the cells showed the positive response for α-SMA and absent response for CK19.Conclusions The hAMSCs can differentiate into ocular surface cells after being induced by hBCFs.And the differentiation mechanism is possibly relevant to mesenchymal cells epithelium.

Objective To compare the intubating conditions between dexmedetomidine and remifentanil when combined with sevoflurane-nitrous oxide (N2O) for anesthesia induction in the pediatric patients.Methods A total of 122 pediatric patients,aged 4-10 yr,of American Society of Anesthesiologists physical status Ⅰ,undergoing elective plastic surgery,were randomly divided into dexmedetomidine group (group D,n =61) and remifentanil group (group R,n=61).Eight percent sevoflurane and 60％ N2O were inhaled for induction of anesthesia,and the fresh gas flow was set at 6 L/min.After disappearance of eyelash reflex,dexmedetomidine 1 μg/kg and remifentanil 1 μg/kg were intravenously injected over 50-60 s in D and R groups,respectively,and 1 min later tracheal intubation was performed.The intubating conditions were graded,and the satisfactory intubating conditions and successful intubation were recorded.The development of adverse cardiovascular reactions and complications such as hyoxemia and laryngospasm before and after intubation and postoperative pharyngodynia was recorded.Results Compared with group D,no significant change was found in the success rate of intubation,rate of satisfactory intubation,intubating condition grade or incidence of postoperative pharyngodynia (P＞ 0.05),and the incidence of hypertension and sinus tachycardia after intubation was significantly increased in group R (P＜0.05).No pediatric patients developed hyoxemia,laryngospasn or sinus tachycardia in two groups.Conclusion When 8％ sevoflurane and 60％ N2O are inhaled for anesthesia induction,combing with dexmedetomidine 1 μg/kg produces better clinical efficacy than combing with remifentanil 1 μg/kg in improving the intubating conditions for pediatric patients.

OBJECTIVETo explore the intersection and regulation mechanism of "efficacy-toxicity network" of Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma and Aconiti Lateralis Radix Praeparata's action gene in the combination environment of Sini decoction with the network pharmacological method.

METHODThe gene interaction network of Aconiti Lateralis Radix Praeparata, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma were mined and established with Cytoscape software and Agilent literature search plug-in. The "efficiency-toxicity network" intersection of Aconiti Lateralis Radix Praeparata was formed according to its effects in anti-heart failure, neurotoxicity and cardiotoxicity. The target genes were clustered with Clusterviz plug-in. And the possible pathways of the "efficacy-tox- icity network" intersection of Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma and Aconiti Lateralis Radix Praeparata were forecasted in DAVID database.

RESULTThere were five genes related to neurotoxicity, cardiotoxicity and anti-heart failure function of Aconiti Lateralis Radix Praeparata, namely AKT1, BAX, HCC, IL6 and IL8, which formed 47 nodes genes in the "efficiency-toxicity network" intersection of Aconiti Lateralis Radix Praeparata. There were 29 and 27 coincident genes in the "efficiency-toxicity network" of Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma and Aconiti Lateralis Radix Praeparata. There were 23 and 17 possible regulatory pathways.

CONCLUSIONIn the combination environment of Sini decoction, Glycyrrhizae Radix et Rhizoma and Zingiberis Rhizoma may regulate the efficiency-toxicity network of Aconiti Lateralis Radix Praeparata by influencing immune-inflammatory signaling pathway, apoptosis-autophagy signaling pathway, nerve cell and myocardial ischemia and hypoxia protection signaling pathways.

Aconitum ; chemistry ; toxicity ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Gene Regulatory Networks ; drug effects ; Humans ; Rhizome ; chemistry ; toxicity

OBJECTIVETo report the results of clinical characteristics, enzyme activity determination and mutation analysis of GLB1 gene in a Chinese patient with mucopolysaccharidosis (MPS) type IVB (Morquio B disease).

METHODA 14-year-old Chinese boy with MPS type IVB was firstly diagnosed by blood leucocytes galactosamine-6-sulfate sulfatase (GALNS) and β-galactosidase (GLB1) determination, who was characterized by short stature, multiplex skeletal abnormalities, difficulty in walking. PCR-sequencing analysis was applied to detect the mutations in GLB1 of the patient.

RESULTThe patient was characterized by dwarfism, pectus carinatum, kyphosis, normal intelligence, and no neurologic damage of spasms, linguistic capacity and so on. The patient had normal GALNS enzyme activity and very low GLB1 enzyme activity [5.03 nmol/(h·mg) vs. normal value 118 - 413 nmol/(h·mg) ] in leukocytes. A compound heterozygous missense mutations c.442C > T(p.R148C)/c.1454A > G(p.Y485C) in GLB1 gene were detected in this patient. The mutation p.Y485C is a novel variant. With the method of gene analysis of new variant, the mutation p.Y485C was considered to be a pathogenic mutation.

CONCLUSIONThe MPS IVB patient showed severe multiple skeletal deformities, normal intelligence, no neurologic damage and very low GLB1 enzyme activity, who carries compound heterozygous mutations p.R148C/p.Y485C. The mutation p.Y485C in GLB1 gene may be a novel pathologic mutation of MPS type IVB.

Adolescent ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Chondroitinsulfatases ; genetics ; metabolism ; DNA Mutational Analysis ; Humans ; Joints ; pathology ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis IV ; enzymology ; genetics ; pathology ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction ; Radiography ; Spine ; diagnostic imaging ; pathology ; beta-Galactosidase ; genetics ; metabolism

A sensitive high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method was established for the determination of eplerenone (EP) in human plasma. The plasma samples of EP were extracted with ethyl acetate and separated by HPLC on a reversed phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate water solution-methanol (30 : 70, v/v). EP was determined with electrospray ionization-mass spectrometry (ESI-MS) in the selected ion monitoring (SIM) mode. The calibration curves were linear over the range of 2-4 000 ng x mL(-1) for EP. The lower limit of quantification was 2 ng x mL(-1). The method has been successfully applied in the pharmacokinetic study of the EP tablets. The main pharmacokinetic parameters of EP after oral administration of 25 mg, 50 mg, 100 mg were as follows, t1/2: (4.9 +/- 2.1), (4.7 +/- 1.5), (5.9 +/- 1.2) h; AUC(0-infinity): (4 402 +/- 1 735), (8 150 +/- 2 509), (13 783 +/- 4 102) microg x h x L(-1); and MRT: (6.2 +/- 2.1), (6.6 +/- 1.3), and (7.2 +/- 1.6) h. Parameters of EP after oral administration of multiple doses of 50 mg were as follows, t1/2: (6.1 +/- 1.7) h; AUC(ss): (10 071 +/- 4220) microg x h x L(-1); MRT: (8.1 +/- 2.3) h; and DF: (3.2 +/- 1.0).
Chromatography, High Pressure Liquid ; methods ; Humans ; Spectrometry, Mass, Electrospray Ionization ; methods ; Spironolactone ; analogs & derivatives ; blood ; pharmacokinetics

OBJECTIVETo observe the acute hemodynamic effects of intravenous diltiazem in patients with congenital heart defect (CHD) and severe pulmonary hypertension (HP) post cardiac surgery.

METHODSFrom November 2003 to September 2005, 12 patients with CHD and severe HP (4 male, mean age 17.8 +/- 9.8 years) after cardiac surgery received intravenous diltiazem (3 - 5 microg x kg(-1) x min(-1)) in the Intensive Care Unit. Mean pulmonary artery pressure (mPAP), mean arterial pressure (MAP), heart rate (HR), stroke volume (SV), systemic vascular resistance (SVR), pulmonary vascular resistance (PVR) were monitored with Swan-Ganz catheter before (T1) and 6 hours (T2) after diltiazem injection, before weaning patients off of ventilator (T3), 1 hour (T4) and 24 hour (T5) after extubation.

RESULTSAll patients survived during the observation period and no patient developed pulmonary hypertension crisis. The average ventilation time was (88.7 +/- 50.1) hours. Mean ICU stay time was (5.8 +/- 3.1) days. Compared to T1, mPAP was significantly decreased at T3 and T5, MAP significantly increased at T4 and T5, HR significantly reduced at T2 and thereafter, SV significantly increased at T3, T4 and T5 and PVR significantly increased at T3 and T5 while SVR remained unchanged after diltiazem therapy.

CONCLUSIONIntravenous use of diltiazem is safe and effective for patients with CHD with severe HP post cardiac surgery.

Adolescent ; Adult ; Child ; Child, Preschool ; Diltiazem ; therapeutic use ; Female ; Heart Defects, Congenital ; complications ; drug therapy ; Humans ; Hypertension, Pulmonary ; complications ; drug therapy ; Injections, Intravenous ; Male ; Young Adult

OBJECTIVEMany children were found to have low free carnitine level in blood by tandem mass spectrometry technology. In some of the cases the problems occurred secondary to malnutrition, organic acidemia and other fatty acid oxidation metabolic diseases, and some of cases had primary carnitine deficiency (PCD). In the present article, we discuss the diagnosis of PCD and evaluate the efficacy of carnitine in the treatment of PCD.

METHODWe measured the free carnitine (C0) and acylcarnitine levels in the blood of 270 000 neonates from newborns screening program and 12 000 children with suspected clinical inherited metabolic diseases by tandem mass spectrometry. The mutations of carnitine transporter protein were tested to the children with low C0 level and the diagnosis was made. The children with PCD were treated with 100 - 300 mg/kg of carnitine.

RESULTSeventeen children were diagnosed with PCD, 6 from newborn screening program and 11 from clinical patients. Mutations were found in all of them. The average C0 level [(2.9 ± 2.0) µmol/L] in patients was lower than the reference value (10 µmol/L), along with decreased level of different acylcarnitines. The clinical manifestations were diverse. For the 6 patients from newborn screening, 4 were asymptomatic, 1 showed hypoglycaemia and 1 showed movement intolerance from 2 years of age. For the 11 clinical patients, 8 showed hepatomegaly, 7 showed myasthenia, 6 showed cardiomyopathy, 1 showed chronic abdominal pain, and 1 showed restlessness and learning difficulty. Among these patients, 14 cases were treated with carnitine. Their clinical symptoms disappeared 1 to 3 months later. The C0 level in the blood rose to normal, with the average from (4.0 ± 2.7) µmol/L to (20.6 ± 8.3) µmol/L (P < 0.01). However, the level was still lower than the average level of healthy children [(27.1 ± 4.5) µmol/L, P < 0.01].

CONCLUSIONSeventeen patients were diagnosed with PCD by the test levels of free carnitine and acylcarnitines in blood with tandem mass spectrometry, and gene mutation test. Large dose of carnitine had a good effect in treatment of the PCD patients.

Cardiomyopathies ; diagnosis ; drug therapy ; genetics ; Carnitine ; analogs & derivatives ; blood ; deficiency ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Female ; Follow-Up Studies ; Humans ; Hyperammonemia ; diagnosis ; drug therapy ; genetics ; Infant ; Infant, Newborn ; Male ; Muscular Diseases ; diagnosis ; drug therapy ; genetics ; Mutation ; Neonatal Screening ; methods ; Organic Cation Transport Proteins ; deficiency ; genetics ; Reference Values ; Tandem Mass Spectrometry