Objective　To explore the effects of hypoxia on the syntheses and secretion of adrenomedullin (AM), calcitonin gene related peptide (CGRP) and c-type natriuretic peptide (CNP) and the relationship between these peptides. Methods　Rat models were established with hypoxia for 10, 20 and 30 d respectively and rats under normal altitude were served as control. Pulmonary artery pressure and the maximum increasing speed of right ventricle (RVdp/dtmax) were measured in every group. The dynamic changes of AM, CGRP and CNP concentrations in plasma were studied with radioimmunoassay. Results　During hypoxia, pulmonary artery pressure and RVdp/dtmax were enhanced. Plasma AM and CNP concentrations were increased while CGRP was decreased significantly. The plasma level of AM had positive correlation with that of CNP, but negatively correlated with that of CGRP. Conclusion　Results indicate that hypoxia may cause pulmonary artery pressure change and right ventricle has compensatory reaction to hypoxic pulmonary hypertension. Dynamic changes of plasma AM, CGRP and CNP concentrations can be regarded as indexes for condition of illness.

Objective To explore the effects of psychological intervention on the mental status and life quality of hepatolenticular degeneration patients during the perioperative period of splenectomy.Methods 64 cases hepatolenticular degeneration patients during the perioperative period of splenectomy were randomly divided into two groups,the intervention group and the control group,each with 32 cases.Both groups received conventional comfortable nursing,and the intervention group had psychological intervention for eight weeks in addition.Symptom checklist (SCL-90) was used to assess their changes of mental health,while the World Health Organization Quality of Life Scale (WHOQOL-100) was used to assess their quality of life before and after psychological intervention.Results Before psychological intervention,there was no statistically siguificant difference between two groups' SCL-90 scores (P ＞ 0.05).After eight weeks of psychological intervention,the sores of somatization,force,interpersonal relationship,depression,anxiety,hostility,paranoid and psychotic factors in SCL-90 of the intervention group were respectively (1.26 ± 0.43),(1.72 ± 0.44),(1.74 ± 0.45),(1.59±0.43),(1.58±0.52),(1.66±0.74),(1.44±0.57),(1.61±0.68),(1.51±0.43),all of which were lower than those of the control group [(1.60 ± 0.49),(2.06 ± 0.50),(1.74 ± 0.45),(1.85 ±0.49),(1.90±0.55),(2.07±0.66),(1.71 ±0.57),(1.95 ±0.64),(1.82 ±0.56)],and the differences were statistically significant (t =2.950,2.888,2.466,2.256,2.386,2.339,2.060,2.484,respectively; P ＞ 0.05).Before psychological intervention,there was also no statistically significant difference between two groups' WHOQOL-100 scores (P ＞ 0.05).And after eight weeks of psychological intervention,the score of every dimension was higher in the intervention group than in the control group,and the differences were statistically significant (t =4.401,4.694,3.242,5.410,4.576,4.847,3.834,respectively;P ＜0.01).Conclusions Psychological intervention can effectively improve the mental health and quality of life of hepatolenticular degeneration patients during the perioperative period of splenectomy,and thus should be strengthened.

The aim of this study was to evaluate the effect of all-trans retinoic acid (ATRA) on cell adhesion molecule expression and adhesion capacity of bone marrow stromal cells (BMSC) in patients after conditioning treatment for peripheral blood stem cell transplantation (PBSCT). BMSC of 27 patients before and after conditioning treatment for PBSCT were cultured in vitro. After treated with ATRA at 0.01, 0.1, or 1 micromol/L, expression of intercellular adhesion molecule-1 (ICAM-1) protein and vascular adhesion molecule-1 (VCAM-1) protein were detected by flow cytometry, and soluble ICAM-1 (sICAM-1) protein was determined by using radioimmunoassay. Then BMSC was co-cultured with CD34+ cells, and adhesion rate of BMSC to CD34+ cells was measured. The results showed that after pretreatment with conditioning regimen for PBSCT, the expressions of ICAM-1 and VCAM-1 proteins in BMSC and the expression level of sICAM-1 protein in supernatant of BMSC culture were down-regulated, and the adhesion rate of BMSC to CD34+ cells was decreased, after administration of ATRA, the expression of ICAM-1 protein in BMSC, sICAM-1 protein in culture medium and adhesion rate of BMSC to CD34+ cells all increased significantly, but expression of VCAM-1 protein changed no significantly. It is concluded that the ATRA can partly restore adhesion function of BMSC injured by pretreatment for PBSCT and contribute to hematopoietic reconstitution.
Adolescent ; Adult ; Antigens, CD34 ; Antineoplastic Agents ; pharmacology ; Bone Marrow Cells ; metabolism ; pathology ; Cell Adhesion ; drug effects ; Cell Adhesion Molecules ; biosynthesis ; genetics ; Child ; Coculture Techniques ; Hematologic Neoplasms ; metabolism ; pathology ; therapy ; Humans ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Stromal Cells ; metabolism ; pathology ; Tretinoin ; pharmacology ; Tumor Cells, Cultured ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

The aim of this study was to evaluate the effect of dimethyl amiloride (DMA), a specific inhibitor of Na(+)/H(+) exchanger-1 (NHE-1), on intracellular pH value (pHi), proliferation and apoptosis of HL-60/ADM cells in vitro. After treatment with DMA at different doses, pHi of HL-60 and HL-60/ADM cell lines were determined by using pH-sensitive fluorescence dye BECEF-AM; the rate of growth inhibition of cells was detected with MTT assay; cell cycle was detected by flow cytometric DNA analysis; cell apoptosis was observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The results showed that pHi in HL-60/ADM cells was higher than that in HL-60 cells. After treatment with DMA at different doses, pHi decreased, the rate of growth inhibition and the rate of apoptotic cells in HL-60/ADM cells were all higher than those in HL-60 cells. Meanwhile, after treatment with DMA during 100 micromol/L to 150 micromol/L, the increase amplitude of G(0)/G(1) phase cells and the decrease amplitude of S + G(2)/M cells in HL-60/ADM cells were higher than those in HL-60 cells. It is concluded that by causing intracellular acidification, the NHE-1-specific inhibitor DMA inhibits proliferation of HL-60/ADM cells and induces apoptosis of HL-60/ADM cells, and the degree of this growth inhibition of HL-60/ADM cells is higher than that of HL-60 cells.
Amiloride ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; HL-60 Cells ; Humans ; Hydrogen-Ion Concentration ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors

In order to study the influence of different gelatin concentrations, and lymphocyte isolation liquid on primary culture of umbilical cord blood-derived adhesive cells (hCBACs), the red blood cells of umbilical cord blood was separated by 3% and 6 % gelatin for detecting the effectiveness of sedimentation, then the adhesion rate at 48 hours, the day of initial expansion and the rate of culture success were detected for hCBACs cultured with CD34(+) cells after the mononuclear cells were separated by 6% gelatin followed by Ficoll and Percoll, and the morphological characteristics and growth status were observed by invert microscopy. Cytochemistry stain for nonspecific esterase stain (NSE), peroxidase (POX), periodic acid Schiff reaction (PAS) and alkali phosphatase (ALP) and immunocytochemistry labeling for CD31, CD45, CD68 and fibronectin (Fn) were detected. The results showed that 6 % gelatin was better than that 3% gelatin for red blood sedimentation. The Percoll was predominant over Ficoll in adhesion rate at 48 hours, the day of initial expansion, the time of initial formation of adhesive cell colony units, the time of maximal numbers of adhesive cell colony units, the the cell fusion time and ratio of culture success. 60% fibroblast-liked cells, 36% macrophage liked cells and 4% small-round cells were observed in cells isolated by both isolated methods. The cytochemistry stain for NSE, POX, PAS and ALP was similar in two groups, the difference was not statistically significant between these two groups. The immunocytochemistry labeling for CD31, CD45, CD68 and Fn was also similar in both groups and the difference was also not statistically significant between these two groups. It is concluded that the combination of 6% gelatin with Percoll is an ideal separation method for primary culture of hCBACs, which provides basic information for clinical application.
Cell Separation ; methods ; Cells, Cultured ; Fetal Blood ; cytology ; Gelatin ; administration & dosage ; pharmacology ; Humans ; Lymphocytes ; cytology

OBJECTIVETo observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells.

METHODSSDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co-cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control.

RESULTSThe level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively.

CONCLUSIONDown-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells.

Bone Marrow Cells ; metabolism ; Cell Adhesion ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; metabolism ; Coculture Techniques ; Drug Resistance, Neoplasm ; Gene Expression ; Humans ; Jurkat Cells ; RNA Interference ; Stromal Cells ; metabolism

OBJECTIVETo study the effects of RNA interference inhibiting stromal cell derived factor-1 (SDF-1) expression on the proliferation and apoptosis of co-cultured Jurkat cells.

METHODInhibition of SDF-1 expression by RNA interference (RNAi) was achieved by transferring SDF-1 specific short hairpin RNA (shRNA) expressing plasmid into cultured human acute leukemic bone marrow stromal cells. Resistant clones were obtained by G418 selection (group A). The concentration of SDF-1 protein in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The population double time (PDT), cell cycles, apoptosis rates and the expressions of PCNA, Bcl-2/Bax, Fas/FasL of co-cultured Jurkat cells were detected by cells counting, flow cytometry. TdT-mediated dUTP nick-end labelling (TUNEL) and immunocytochemistry (ICC), respectively. The un-transfected acute leukemic (group B) and normal (group C) bone marrow stromal cells were taken as controls.

RESULTSThe content of SDF-1 protein in supernatant of group A\[(384 +/- 41) pg/ml] was significantly lower than that in group B[(2474 +/- 271) pg/ml] or group C[(1324 +/- 154) pg/ml]. As group A compared with group B and group C, the PDT of co-cultured Jurkat cells was prolonged (group A: 42 h, vs group B: 29 h, group C: 33 h), and G(0)/G(1) stage cells increased [group A: (28.47 +/- 2.39)%, vs group B: (19.43 +/- 2.80)%, group C: (27.15 +/- 2.07)%], S stage cells decreased [group A: (25.57 +/- 1.90)%, vs group B: (74.48 +/- 3.23)%, group C: (60.99 +/- 2.33)%], G(2)/M stage cells increased [group A: (45.96 +/- 3.24)%, vs group B: (6.09 +/- 1.96)%, group C: (11.86 +/- 1.98)%], the apoptosis rate increased [group A: (15.2 +/- 0.8)%, vs group B: (5.4 +/- 0.7)%, group C: (9.5 +/- 0.4)%], and the expressions of PCNA, Bcl-2, Fas decreased; whereas the expressions of Bax and FasL were increased.

CONCLUSIONThe inhibition of SDF-1 expression in bone marrow stromal cells inhibits the proliferation and promotes the apoptosis of co-cultured Jurkat cells.

Apoptosis ; genetics ; Bone Marrow Cells ; metabolism ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; Coculture Techniques ; Gene Expression ; Humans ; Jurkat Cells ; RNA Interference ; Stromal Cells ; metabolism ; Transfection

Blood Coagulation Disorders ; drug therapy ; Child ; Child, Preschool ; Drug Therapy, Combination ; Female ; Heparin ; administration & dosage ; Humans ; Male ; Nephrotic Syndrome ; blood ; complications ; urine ; Urokinase-Type Plasminogen Activator ; administration & dosage

BACKGROUNDIn China, transthoracic echocardiography (TTE) is popularly used for pre-intervention examination for atrial septal defect (ASD) and for guiding ASD closure. However, the ability to determine ASD size and the safety and efficacy of TTE for guiding ASD closure still has not been widely accepted. This study aimed to evaluate the efficacy and safety of TTE used before, during and after transcatheter ASD closure with Amplatzer septal occluders (ASO).

METHODSSixty-eight subjects (15 men and 53 women; mean age (33.7 +/- 17.3) years) were enrolled. TTE was used to measure the diameters and guide transcatheter closure of ASD. The ASD was examined by long-axis view, basal short-axis view, apical four-chamber view and the subcostal view to observe position, diameter and relation with neighbouring structures. The largest diameter was selected as the reference diameter. Patients were divided into 3 groups according to the ASD reference diameter: 22 subjects with ASD diameter 4 - 14 mm (group A); 21 subjects with ASD diameter 15 - 20 mm (group B); and 25 subjects with ASD diameter 21 - 33 mm (group C).

RESULTSASD was occluded successfully in groups A and B. In group C, occlusion failed in 2 cases; 1 case remained with a 3-mm residual shunt sustained until 6-month follow-up. However, at 6-month follow-up, no case of thromboembolism, ASO dislocation or death occurred in the three groups. The diameter of ASD measured by TTE could accurately predict the ASO size that could successfully occlude the ASD, especially in patients with ASD < 20 mm. The ASD diameter measured by TTE correlated well with ASO size (r = 0.925, P < 0.001; r = 0.976, P < 0.001; r = 0.929, P < 0.001 respectively).

CONCLUSIONSASD diameter measured by TTE can accurately estimate the size of the ASO needed for successful closure of ASD. The larger the ASD, the much larger the ASO needed. TTE is a satisfactory guiding imaging tool for ASD closure.

Adolescent ; Adult ; Aged ; Balloon Occlusion ; instrumentation ; Child ; Child, Preschool ; Echocardiography ; methods ; Female ; Heart Septal Defects, Atrial ; diagnostic imaging ; therapy ; Humans ; Male ; Middle Aged

Objective To investigate hepatocyte growth factor(HGF)gene expression and biological effect after gene transfection into penumbra tissue in rat cerebral ischemic model.Methods Human HGF cDNA was ligated to pIRES2-EGFP vector.The recombinant plasmid was transfected into the penumbra tissue with liposome.Brains of treated and control animals were analyzed 7 days later.Expression of HGF protein was determined by fluorescence microscopy and immunohistochemistry.Vessel numbers were quantified.Changes of cerebral blood flow(CBF)was detected by CT perfusion.Results Enzymatic digestion and electrophoresis confirmed that HGF fragment had been correctly cloned into BamH I and Sal I sites of pIRES2-EGFP.After HGF gene transfection,expression of HGF in transfected neurocytes was observed with fluorescence microscopy and immunohistochemistry.The number of vessels was significantly increased in penumbra tissue transfected with HGF vector as compared with control vector(46.71?7.11, 20.43?3.21,18.00?3.27,respective,F = 74.447;P