Animals ; Asthma ; drug therapy ; metabolism ; Genistein ; pharmacology ; therapeutic use ; Guinea Pigs ; Interleukin-4 ; genetics ; metabolism ; Janus Kinase 1 ; genetics ; metabolism ; Lung ; metabolism ; Male ; Protein Kinase Inhibitors ; pharmacology ; therapeutic use

Background Retinal retinoic acid (RA) plays an important role in the formation of the lensinduced myopia.However,it is not clear how RA transfer the myopic signal to choroid throughout the retinal pigment epithelium (RPE) barrier.Objective The aim of this study was to investigate the effect of all-trans retinoic acid (atRA) on the barrier of RPE in lens-induced myopic eye of guinea pig.Methods Thirty left eyes of 30 21-dayold clean guinea pigs were randomized into normal control group and the model group.The models of out of focus were induced by covering of-6.00 D concave lens on the left eyes for 15 days.Radius of corneal curvature was measured using corneal curvimeter,and diopeter of the guinea pig was examined by mydriatic optometry.The length of ocular axis was detected by A-sonography.The animals were sacrificed and the retinas of the left eyes were isolated for the culture and passage of RPE cells.The third generation of cells were incubated Millcell-PET microporous film,and atRA at the concentration of 1 × 10-6,1 × 10-7,1 × 10-8 and 1 × 10-9 mol/L was added to the micropore respectively for 12 hours,and the micropores with equal-solvent served as negative control group.Methyl thiazolyl tetrazolium (MTT)colorimetric method was used to detect the survival rate of the cells.Subsequently,the transepithelial electrical resistance (TER) of the monolayer cells was determined with CN10-EVOM2 resistance measuring meter.The vesicular transport change of RPE membrane in different groups was evaluated by FM1-43 fluorescence staining.Results The mean diopter was (-2.20±0.95) D in the models,and that in the controls was (+ 1.15 ±0.30) D,with a significant difference between them (t =14.57,P＜ 0.01).The axial length was (8.24 ± 0.09) mm in the models and it was significantly longer than (7.81±0.05) mm in the controls (t=17.20,P＜0.01).RPE cells grew well to form a monolayer in Millcell culture pool after one week.After 24 hours of the atRA treatment,the survival rate of RPE cells reduced gradually with the increase of atRA concentration with the highest rate in the 1 × 10-9 mol/L atRA group (93.3 ％) and followed by the 1 × 10-8 mol/L atRA group (88.2％).More than half of the cells dead in the 1 × 10-6mol/L and 1 × 10-7mol/L atRA groups (53.8％ and 47.1％).Significant differences in the TER value and fluorescence staining intensity of the cells were seen among the various groups (F =43.89,P =0.00 ; F =26.13,P =0.00),with the maximal values in the 1 × 10-8mol/L atRA group.The FM1-43 fluorescence located on the cellular membrane and cytoplasm.Conclusions AtRA can increase the functional state of tight junction and vesicular transport,which regulated the RPE cell barrier in the guinea pig.

In order to reveal the cause of disease occurred in the process of Coptis chinensis growth, this paper studied the bacterial species diversity index of different aged rhizospheric and non-rhizospheric soil planting normal or sick C. chinensis by using PCR-DGGE technique. The representative DGGE bands were chosen to be cloned, and sequenced, the phylogeny were constructed. The results showed that the bacterial communities were very different between the normal and diseased soil samples of C. chinensis, and the diversity index (H) of diseased soil samples were higher than that of normal soil samples. Sequencing analysis of representative cloned DGGE bands showed that the unculturable bacteria were the dominant groups, and bacteria belonged to genus Bacillus, Acidovorax, Acinetobacter, uncultured Kluyvera, and uncultured Comamonas were also existing, but the reported plant pathogenic bacteria were not found in the C. chinensis planting soil. The density and brightness of clone band d in diseased soil samples was higher than that in normal soil sample, and sequencing analysis showed that it belonged to genus Acidovorax. Obviously, during the process of C. chinensis growth, the rhizospheric bacteria population changed, and the quantity of bacteria belong Acidovorax increased, which probably resulted in the disease occurred during C. chinensis growth.
Bacteria ; classification ; genetics ; isolation & purification ; Biodiversity ; Coptis ; growth & development ; microbiology ; Denaturing Gradient Gel Electrophoresis ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Rhizosphere ; Soil Microbiology

OBJECTIVETo explore the molecular mechanism of Qiangxin Fumai granule (QFG, an effective Chinese composite drug) in preventing and treating sick sinus syndrome (SSS).

METHODRabbit model of sinoatrial ischemia/reperfusion was established by occluding and loosening the root of right coronary artery. Effect of QFG on cell apoptosis was observed by TUNEL method, and its effect on apoptotic related gene Bax, Bcl-2 and Fas-L gene protein expression was observed by immunohistochemical method. Average light density values of the expression of Bax, Bcl-2 and Fas-L of SAN cells was determined by Imagepro Plus image analysis system.

RESULTSinoatrial injury induced by ischemia/reperfusion could cause evident sinoatrial cell apoptosis, enhance Fas-L gene protein expression and obviously enhance Bax gene protein expression, reduce Bcl-2/Bax ratio. QFG could significantly down-regulate Fas-L and Bax gene protein expression, up-regulate Bcl-2/Bax ratio, significantly inhibit and block the sinoatrial cell apoptosis.

CONCLUSIONTo inhibit and block the event of cell apoptosis through regulating Bax, Bcl-2 and Fas-L gene protein expression in sinoatrial node after ischemia/reperfusion might be one of the mechanisms of QFG in preventing and treating sinoatrial ischemia/reperfusion injury of SSS.

Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Microscopy, Fluorescence ; Muscle Cells ; cytology ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; Random Allocation ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo evaluate the correlations of microvessel density (MVD), vascular endothelial growth factor (VEGF), thrombospodin1 (TSP1) and p53 protein with prognosis in epithelial ovarian cancer.

METHODSSamples from 57 patients with primary epithelial ovarian cancer were examined by immunohistochemical staining using anti-VEGF, anti-TSP1, anti-p53 and anti-CD34 antibodies. The correlation of MVD, expression of VEGF, TSP1 and p53 protein with postoperative recurrence and overall survival were analyzed retrospectively.

RESULTSVEGF, TSP1 and p53 protein was positively detected in 40 (70.2%), 27 (47.4%) and 35 (61.4%) of those patients, respectively. The mean MVD in this series was 30.3 +/- 8.5. High MVD, positive VEGF expression and negative TSP1 expression were positively correlated with postoperative recurrence. Univariate analysis showed that patients with high MVD, positive expression of VEGF and p53 had shorter median overall survival time than those with lower MVD, negative expression of VEGF and p53 (P = 0.0187, P = 0.010 and P = 0.005, respectively), while TSP1 expression was revealed as a protective factor for prognosis. Patients with positive expression of TSP1 had longer median overall survival time than those with negative TSP1 expression (P = 0.042). Multivariate analysis showed that MVD and p53 expression were two independent prognostic factors in epithelial ovarian cancer (P = 0.018 and P = 0.009, respectively).

CONCLUSIONVEGF, TSP1 and p53 protein may play an important role in the angiogenesis of epithelial ovarian cancer. High MVD level and p53 protein expression are two independent poor prognostic factors.

Adenocarcinoma, Clear Cell ; blood supply ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Cystadenocarcinoma, Serous ; blood supply ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Microvessels ; pathology ; Middle Aged ; Multivariate Analysis ; Neoplasm Recurrence, Local ; Ovarian Neoplasms ; blood supply ; metabolism ; pathology ; Retrospective Studies ; Survival Rate ; Thrombospondin 1 ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the ECG and electrophysiological characteristic of patients with idiopathic ventricular tachycardia (VT) and premature ventricular contraction (PVC) originating from left (LVOT) and right (RVOT) ventricular outflow tracts and assess the clinical effect of radio frequency catheter ablation (RFCA) on these patients.

METHODSRFCA was performed in 58 patients (10 with VT and 48 with PVC, 5 patients with VT from RVOT under the guidance of non-contact mapping system Ensite3000). VT or PVC originated from LVOT in 15 patients (12 out of 15 from left sinus of Valsalva) and RVOT in 43 patients.

RESULTS(1) R wave in II, III, aVF leads was the common characteristics of VT or PVC originated from LVOT and RVOT and difference in wave duration index and R/S-wave amplitude ratio in V(1) or V(2) could be used to define VT and PVC originated from LVOT or RVOT. (2) Ablation was successful in 55 out of 58 patients (9 patients with the 2nd ablation, evaluated as arrhythmia-free at 3 months post ablation without medication) and failed in 3 patients. One patient developed pericardial tamponade during ablation and recovered without complication after related treatments.

CONCLUSIONSRFCA is an effective, safe and curative therapy for VT or PVC originated from LVOT and RVOT. Non-contact mapping system (Ensite3000) is a safe and reliable tool to guide mapping and ablation in patients with complex VT and unstable hemodynamics.

Adolescent ; Adult ; Aged ; Catheter Ablation ; Female ; Humans ; Male ; Middle Aged ; Tachycardia, Ventricular ; etiology ; therapy ; Ventricular Outflow Obstruction ; complications ; Ventricular Premature Complexes ; etiology ; therapy ; Young Adult

OBJECTIVEGerm cells are much more susceptible to oxidative stress than somatic cells. Accumulating evidence indicates that uncoupling protein 2 (UCP2) prevents the apoptosis of multiple types of cells induced by reactive oxygen species (ROS). The present study aimed to evaluate the protective effect of uncoupling protein 2 (UCP2) against oxidative damage to human sperm.

METHODSSemen parameters were obtained from 97 semen samples by computer assisted sperm analysis (CASA), and the samples, based on the percentage of grade a + b sperm, allotted to Groups I (n = 25, grade a + b sperm > 50%), II (n = 24, grade a + b sperm 25%-50%), III (n = 24, grade a + b sperm 10-25%) and IV (n = 24, grade a + b sperm < 10%). The expressions of sperm UCP2 mRNA was detected by RT-PCR, and the malonaldehyde (MDA) content in ROS was determined in the sperm suspension.

RESULTSThe expressions of UCP2 mRNA in sperm were 1.51 +/- 0.24, 1.28 +/- 0.15, 1.17 +/- 0.20 and 0.69 +/- 0.18, and the MDA contents were (14.66 +/- 2.55), (16.00 +/- 2.09), (17.44 +/- 1.40) and (21.20 +/- 3.50) nmol/10(8) sperm in Groups I, II, III and lV, respectively. There was a significant negative correlation between the expression of UCP2 mRNA and the content of MDA (r = -0.633, P < 0.01), the former obviously reduced while the latter remarkably increased in the groups of lower sperm motility (P < 0.01).

CONCLUSIONUCP2 plays an important protective role against oxidative stress damage to human sperm by diminishing ROS production.

Adult ; Humans ; Ion Channels ; metabolism ; Male ; Middle Aged ; Mitochondrial Proteins ; metabolism ; Oxidative Stress ; Reactive Oxygen Species ; metabolism ; Sperm Count ; Sperm Motility ; Spermatozoa ; metabolism ; Uncoupling Protein 2

OBJECTIVETo study the clinical efficacy of porcine pulmonary surfactant (PS) combined with budesonide suspension intratracheal instillation in the treatment of neonatal meconium aspiration syndrome (MAS).

METHODSSeventy neonates with MAS were enrolled for a prospective study. The neonates were randomly assigned to PS alone treatment group and PS+budesonide treatment group (n=35 each). The PS alone treatment group was given PS (100 mg/kg) by intratracheal instillation. The treatment group was given budesonide suspension (0.25 mg/kg) combined with PS (100 mg/kg).

RESULTSThe rate of repeated use of PS in the PS+ budesonide group was significantly lower than that in the PS alone group 12 hours after treatment (p<0.05). The improvement of PaO/FiO, TcSaO, PaO, and PaCOin the PS+ budesonide group was significantly greater than that in the PS alone group 6, 12, and 24 hours after treatment (p<0.05). The chest X-ray examination showed that the pulmonary inflammation absorption in the PS+ budesonide group was significantly better than that in the PS alone group 48 hours after treatment (p<0.05). The incidence of complications in the PS+budesonide group was significantly lower than that in the PS alone group (p<0.05), and the average hospitalization duration was significantly shorter than that in the PS alone group (p<0.01).

CONCLUSIONSPS combined with budesonide suspension intratracheal instillation for the treatment of neonatal MAS is effective and superior to PS alone treatment.

Animals ; Budesonide ; administration & dosage ; Female ; Humans ; Infant, Newborn ; Length of Stay ; Male ; Meconium Aspiration Syndrome ; complications ; drug therapy ; Prospective Studies ; Pulmonary Surfactants ; administration & dosage ; Suspensions ; Swine ; Trachea

This article reported the clinical manifestations, steroid profiles and adrenal ultrasound findings in two unrelated Chinese girls with lipoid congenital adrenal hyperplasia (LCAH). Direct DNA sequencing and restriction fragment length polymorphism (RFLP) analysis were used to identify the mutations of steroidogenic acute regulatory protein (StAR) gene. The two patients with 46,XX karyotype, presented hyperpigmentation, growth retardation, and hyponatremia. Steroid profiles analysis revealed elevated plasma adrenocorticotrophic hormone levels, decreased or normal serum cortisol levels and low levels of androgens. Ultrasound examinations revealed that enlarged adrenals in patient 1 and normal adrenals in patient 2. Direct DNA sequencing of StAR gene showed a reported homozygous for c.772C>T(p.Q258X) in patient 1. Compound heterozygous for c.367G>A(p.E123K) and IVS4+2T>A (both novel mutations) were found in patient 2, inherited from her mother and father respectively. The amino acid of mutant position of the novel p.E123K was highly conserved in ten different species and was predicted to have impacts on the structure and function of StAR protein by the PolyPhen-2 prediction software. RFLP analysis revealed three bands (670, 423 and 247 bp) in patient 2 and her father and two bands (423 and 247 bp) in her mother and 50 controls. It is concluded that LCAH should be considered in girls with early onset of adrenal insufficiency and that steroid profiles, karyotype analysis, adrenal ultrasound and StAR gene analysis may be helpful for the definite diagnosis of LCAH.
46, XY Disorders of Sex Development ; diagnosis ; genetics ; Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Amino Acid Sequence ; Child ; Female ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Mutation ; Phosphoproteins ; genetics ; Polymorphism, Restriction Fragment Length

OBJECTIVETo investigate the relationship between the expression of uncoupling protein 2 (UCP2) and ratio of interleukin-10/interferon-gamma (IL10/IFNgamma) in the macrophages from patients with unexplained recurrent spontaneous abortion (URSA).

METHODSTwelve women undergoing selective termination of normal early pregnancy (control) and 11 having URSA were included in this study. Magnetic cell sorting (MACS) was used to isolate the macrophages in the decidua, and the expression of UCP2 was detected with flow cytometry. Cytokine (IL10 and IFNgamma) secretion by the macrophages was detected by enzyme-linked immunosorbent spot-forming (ELISPOT) cell assay.

RESULTSCompared to the control group, the women with URSA showed significantly decreased expression of UCP2 on decidual macrophage (136-/+25 vs 201-/+31, P<0.01), and the expression of UCP2 was positively correlated to the ratio of IL10/IFNgamma(r=0.73, P<0.01).

CONCLUSIONUCP2 may play an important role in the regulation of macrophage activity and cytokine secretion to contribute to spontaneous abortion.

Abortion, Habitual ; metabolism ; Adult ; Cytokines ; metabolism ; Decidua ; cytology ; metabolism ; Female ; Humans ; Interferon-gamma ; secretion ; Interleukin-10 ; secretion ; Ion Channels ; metabolism ; Macrophage Activation ; Mitochondrial Proteins ; metabolism ; Pregnancy ; Uncoupling Protein 2