Aim To investigate subcellular distribution and accumulation of ADR in the sensitive and multidrug-resistant HL-60 cells and its relation to multidrug resistance.Methods The subcellular distribution and accumulation of ADR were studied by confocal scanning laser microscope and flow cytometry.The effects of verapamil,BSO,brefeldin A and chloroquine on ADR distribution and accumulation in HL-60/ADR cells were also examined.Rhodamine123,NBD-ceramide and neutral red were used as fluorescent probes to stain the mitochondria,Golgi apparatus and lysosomes respectively were used to identify the subcellular compartments where ADR was sequestered.Results In drug-sensitive cell line HL-60,ADR fluorescence distributed evenly in the nucleus and cytoplasm,while in multidrug-resistant cell line HL-60/ADR,ADR fluorescence distributed in a punctated pattern in the cytoplasm and was reduced in the nucleus.The mode of ADR distribution in HL-60/ADR cells is highly similar to that of NBD-ceramide.BSO and brefeldin A,instead of verapamil and chloroquine could reverse the abnormal distribution and accumulation of ADR in HL-60/ADR cells.Conclusions The change of ADR distribution and reduction of ADR accumulation in multidrug-resistant cell line was involved in the mechanism of multidrug resistance.

Relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains a main question on treatment failure.Current strategies for management that usually include salvage chemotherapy,donor lymphocytic infusion and second transplantation.Our study assessed the efficacy of decitabine (DAC) for treating patients with acute lymphoblastic leukemia (ALL) who relapsed after allogeneic hematopoietic stem cell transplantation (allo-HSCT).We retrospectively analyzed the outcomes of 12 patients with relapsed ALL after allo-HSCT who received DAC therapy.Nine patients received DAC combined with chemotherapy and donor stem cell infusion,and 3 patients received single-agent DAC.Ten of the 12 patients achieved complete remission (CR),1 achieved a partial remission (PR),and 1 had no response (NR) after treatment at the latest follow-up (LFU),the median survival was 11.2 months (range,3.8-34,7 months).The 1-and 2-year overall survival (OS) rates were 50％ (6/12) and 25％ (3/12),respectively.Five patients were still alive;4 had maintained CR and 1 was alive with disease.Patients with Philadelphia chromosome-positive ALL had higher survival rate than patients with Philadelphia chromosome-negative ALL (57.1％ vs.20％).No aggravated flares of graft-versus-host disease (GVHD) were observed during DAC treatment.Therefore,DAC may be a promising therapeutic agent for ALL recurrence after allo-HSCT.

OBJECTIVETo investigate the mechanical characteristics of new anatomic plate of distal tibia from view of biomechanics.

METHODSTwelve fresh adult moist ankle specimens were randomly divided into four block groups (every group had 3 specimens), 3 tibial specimens as a normal control (normal group N), 9 specimens were resulted in unstable distal tibial Pilon fracture. Using steel plate fixation with a new anatomic distal tibial plate (group A), reconstruction plate (group B), clover plate (group C). Group B and group C as control group to test the remote axial compressive strength, remote axial stiffness, reversing biomechanical properties, contacted characteristics of the tibial astragaloid articular surface.

RESULTSThe remote axial compressive strength, remote axial stiffness, reversing biomechanical properties, contacted characteristics of tibial astragaloid articular surface the in distal tibial Pilon fracture instability of group A were near normal group N (P>0.05). Group A was best than group B and C (P<0.05).

CONCLUSIONThe new anatomic plate of distal tibia was relatively strong, which can reach effective and stable fixation for unstable distal tibial Pilon fracture.

Biomechanical Phenomena ; Bone Plates ; Fracture Fixation, Internal ; Humans ; In Vitro Techniques ; Tibia ; chemistry ; injuries ; surgery ; Tibial Fractures ; surgery

OBJECTIVETo present the technique and experience of robotic-assisted laparoscopic radical cystectomy (RARC) by da Vinci surgical system.

METHODSFrom December 2007 to September 2008, 4 patients underwent RARC and urinary diversion. The age of patients was 44 to 63 years old. The body mass index was 22.8 to 27.7. All their clinical stages were lower than T2N0M0. The technique for RARC involving ureters dissection, posterior dissection, lateral pedicle control, anterior dissection, dorsal vein complex control, neurovascular bundles sparing, lymphadenectomy, ureter-ileal anastomosis, urethra-neobladder anastomosis to either ileal conduit urinary diversion or neobladder reconstruction performed extracorporeally.

RESULTSAll the operations were accomplished successfully. The urinary diversion of 2 case was ileal conduit and others was ileal orthophoria neobladder. The operation time was 300 to 450 min. The time of radical cystectomy was 150 to 180 min. The estimated blood loss was 100 to 500 ml. The postoperative hospital stay was 9 to 35 d. The bed rest time was 4 to 9 d. There was 1 patients who had incomplete intestinal obstruction at 8th postoperative day cured by conservative therapy. The patients were followed up for 3 to 12 months, all patients survived without tumor recurrence. The patients have satisfied urinary continence and normal renal functions without hydronephrosis after the operation.

CONCLUSIONSRARC is small incision and safe, the results are definite. It is one of the direction of minimally invasive urologic surgery.

Adult ; Cystectomy ; methods ; Follow-Up Studies ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Robotics ; Urinary Diversion ; methods

OBJECTIVETo study the in vitro effects of low-molecular weight heparin (LMWH) and dexamethasone on the hemolysis of red blood cells from paroxysmal nocturnal hemoglobinuria (PNH) patients.

METHODSBy Ham's test and micro-complement lysis sensitive test (mCLST), the changes of hemolysis of red blood cells from 6 PNH patients were tested by adding different doses of LMWH and dexamethasone into the test mixture. The effects of LMWH and dexamethasone on the coagulation of the tested blood samples were also studied by activated partial thromboplastin time (APTT).

RESULTS(1) Either LMWH or dexamethasone could dose-dependently inhibit the hemolysis of PNH red blood cells, and the effects were synergistic when added together. The same dose of LMWH induced a less than 100% prolongation of APTT. (2) Dexamethasone could inhibit the hemolysis in Ham's test and had different effects on the hemolysis by different adding methods in mCLST. LMWH could inhibit the hemolysis in both Ham's test and mCLST.

CONCLUSIONBoth LMWH and dexamethasone could inhibit the hemolysis of PNH red cells and showed a synergistic effect. The mechanisms of the inhibition of hemolysis were different. Furthermore, a tolerable dose of LMWH induced only a limited prolongation of APTT, which might be useful for controlling acute hemolysis and reducing the dose of dexamethasone.

Anti-Inflammatory Agents ; pharmacology ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Erythrocytes ; cytology ; drug effects ; Hemoglobinuria, Paroxysmal ; blood ; Hemolysis ; drug effects ; Heparin, Low-Molecular-Weight ; pharmacology ; Humans ; Partial Thromboplastin Time

OBJECTIVETo summarize the clinical characteristics and outcome of patients with long-QT syndrome (LQTs) accompanied with torsade de pointes.

METHODSThirty-two eligible patients were included in this study. Clinical and electrocardiographic data were analyzed and telephone or out-patient follow-up were made in all patients.

RESULTSThere were 15 patients with inherited LQTs (h-LQTs) and 17 patients with acquired LQTs (a-LQTs). There are more women (n = 24) than men (n = 8). β blockers, potassium and magnesium supplement were the basic therapy for h-LQTs patients, bivent pacemaker was implanted in 2 patients and implantable cardioverter defibrillator was implanted in 5 patients. Ventricular tachyarrhythmias and syncope occurred in 4 patients during (39.4 ± 25.1) months follow-up. In 17 a-LQTs patients, one patient with dilated cardiomyopathy died suddenly and another patient with implanted cardioverter defibrillator experienced one ventricular tachycardia during (30.9 ± 13.3) months follow-up.

CONCLUSIONSThe prognosis in h-LQTs and a-LQTs patients with structure heart disease is poor. ICD or CRT-D therapy is suggestive for a-LQTs patients with structure heart disease.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Long QT Syndrome ; complications ; therapy ; Male ; Middle Aged ; Pacemaker, Artificial ; Torsades de Pointes ; complications ; therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the mechanism of acupuncture for treatment of asthma.

METHODSForty SD rats were randomly divided into 5 groups: blank control group, normal saline control group (NS control group), asthma model group, asthma model with acupuncture group (asthma acupuncture group) and asthma model with binding group (asthma binding group). The asthma acupuncture group was treated with acupuncture at "Dazhui" (GV 14), "Feishu" (BL 13) and "Fengmen" (BL 12); the asthma binding group was only binding without acupuncture and no intervention was given in the other groups. Surfactant protein-A (SP-A) expression was examined by Western-Blot.

RESULTSAirway resistance in asthma model group was significantly higher than that in blank control group and NS control group from 3 to 6 min after asthma provocation (all P<0.01). Western-Blot detection showed that SP-A expression in bronchoalveolar lavage fluid (BALF) of the rats in asthma model group was significantly lower than that in blank control group and NS control group (both P<0.05), and which in asthma acupuncture group was significantly higher than that in asthma model group (P<0.05).

CONCLUSIONThe mechanism of prevention and treatment of acupuncture for allergic asthma is related to regulating SP-A expression of airway in asthmatic rats.

Acupuncture Therapy ; Animals ; Asthma ; genetics ; immunology ; therapy ; Bronchoalveolar Lavage Fluid ; immunology ; Disease Models, Animal ; Gene Expression ; Humans ; Male ; Pulmonary Surfactant-Associated Protein A ; genetics ; immunology ; Random Allocation ; Rats ; Rats, Sprague-Dawley

To construct gene vaccine of PPV and to investigate the effects of interleukin 2 (IL-2) as an adjuvant on immune responses in mouse, the recombinant expression plasmid of pCIneo-IL2-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluore assay. To study the immune effects of DNA vaccine in vitro and in vivo, mice were used as the animal model. The recombinant plasmid pCIneo-IL2-VP2, the control plasmid pCI-neo and the PPV live vaccine were immunized by intramuscular injection. Anti-PPV antibodies were measured by ELISA, lymphocyte proliferation activity was detected using MTT method, and the specific killing activities of CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to highest after four weeks. Compared with the control group, the pCIneo-IL2-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-IL2-VP2 induced humoral and cellular immunity responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immunity response in mice with the IL-2 gene as an adjuvant.
Adjuvants, Immunologic ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunization ; Interleukin-2 ; genetics ; immunology ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

Objective:To explore the targets and relevant signaling pathways of Suoquanwan in the treatment of enuresis using network pharmacology，and animal expriments are applied to further define its mechanism of action. Method:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP） database was used to screen out active chemical components of Suoquanwan，varieties of systematic biological databases were integrated to construct the "active component-disease-target" network relationship，and the common protein protein interaction network（PPI） network genes were functionally enriched. Quantitative real time polymerase chain reaction（Real-time PCR） and Western blot were used to verify the effect of Suoquanwan on AVPR2 and DRD2 gene. Result:A total of 32 active ingredients were screened from Suoquanwan. These active ingredients were interacted with 131 potential targets relating to Enuresis，which contained 14 core target genes，namely arginine vasopressin receptor 2 （AVPR2）， neurotrophic receptor tyrosine kinase 1（NTRK1）， dopamine receptor D2（DRD2）， opioid receptor mu 1（OPRM1）， 5-hydroxytryptamine receptor 1A（HTR1A）， 5-hydroxytryptamine receptor 1B（HTR1B），solute carrier family 6 member 4（SLC6A4），Adrenoceptor Alpha 2A（ADRA2A）， prostaglandin-endoperoxide synthase 2（PTGS2）， cholinergic receptor muscarinic 2（CHRM2）， solute carrier family 6 member 3 （SLC6A3）， 5-hydroxytryptamine receptor 6（HTR6）， solute carrier family 6 member 2（SLC6A2）， cytochrome P450 family 2 subfamily C member 19（CYP2C19）. Gene enrichments mainly involved to G protein-coupled receptor signaling pathway，regulation of trans-synaptic signaling，regulation of neurotransmitter transport and neuroactive ligand-receptor interaction. Real-time PCR and Western blot results showed that Suoquanwan could enhance the expression of AVPR2 in rat kidney，and weaken the expression of DRD2 in rat adrenal. Conclusion:The main chemical constituents in Suoquanwan may alleviate enuresis by regulating AVPR2 and DRD2 and then participating in the G protein-coupled receptor signaling pathway，regulation of trans-synaptic signaling，regulation of neurotransmitter transport and other biological processes.

OBJECTIVETo investigate the clinical features, diagnosis and treatment of glucose transporter 1 deficiency syndrome (GLUT1-DS), as well as the diagnostic value of movement disorders.

METHODSThe clinical data of four children with GLUT1-DS were collected, and their clinical features, treatment, and follow-up results were analyzed.

RESULTSThere were two boys and two girls, with an age of onset of 2-15 months. Clinical manifestations included movement disorders, seizures, and developmental retardation. Seizures were the cause of the first consultation in all cases. The four children all had persistent ataxia, dystonia, and dysarthria; two had persistent tremor, two had paroxysmal limb paralysis, and two had eye movement disorders. Paroxysmal symptoms tended to occur in fatigue state. All four children had reductions in the level of cerebrospinal fluid glucose and its ratio to blood glucose, as well as SLC2A1 gene mutations. The four children were given a ketogenic diet, at a ketogenic ratio of 2:1 to 3:1, and achieved complete remission of paroxysmal symptoms within 5 weeks.

CONCLUSIONSGLUT1-DS should be considered for epileptic children with mental retardation and motor developmental delay complicated by various types of movement disorders. The ketogenic diet is effective at a ketogenic ratio of 2:1 to 3:1 for the treatment of GLUT1-DS.

Carbohydrate Metabolism, Inborn Errors ; diagnosis ; genetics ; therapy ; Child ; Child, Preschool ; Female ; Humans ; Male ; Monosaccharide Transport Proteins ; deficiency ; genetics ; Movement Disorders ; diagnosis ; genetics ; therapy