AIMTo study the effects of hydrocortisone sodium succinate on sodium current in human atrial myocytes and in guinea pig ventricular myocytes.

METHODSSingle cardiac myocytes were isolated by enzyme. The effects of hydrocortisone sodium succinate on sodium current (INa) were assessed by applying whole-cell patch clamp techniques.

RESULTSHydrocortisone sodium succinate (1, 3, 10 micromol x L(-1)) was shown to inhibit INa of both human atrial myocytes and guinea pig ventricular myocytes in concentration dependent manner and the IC50 were 6.97 and 8.74 micromol x L(-1), respectively. The inhibition effects acted quickly (1-3 min) and the maximal activating voltage of INa was not changed in both human and guinea pig cardiac myocytes.

CONCLUSIONHydrocortisone sodium succinate can exhibit inhibitory effects on INa in both human and guinea pig cardiac myocytes, and its inhibitory effects act rapidly, which are not consistent with genomic effects, so there may be nongenomic effects.

Adolescent ; Adult ; Animals ; Cell Separation ; Child ; Child, Preschool ; Guinea Pigs ; Heart Atria ; pathology ; Heart Defects, Congenital ; pathology ; Heart Ventricles ; cytology ; Humans ; Hydrocortisone ; analogs & derivatives ; pharmacology ; Myocytes, Cardiac ; drug effects ; physiology ; Patch-Clamp Techniques ; Sodium Channels ; drug effects

BACKGROUNDPancreatic islet cell transplantation is an effective approach to treat type 1 diabetes. However, this therapy is not widely used because of the severe shortage of transplantable donor islets. This study investigated whether mesenchymal stem cells (MSCs) derived from human umbilical cord blood (UCB) could be transdifferentiated into insulin producing cells in vitro and the role of extracellular matrix (ECM) gel in this procedure.

METHODSHuman UCB samples were collected and MSCs were isolated. MSCs specific marker proteins were analyzed by a flow cytometer. The capacities of osteoblast and adipocyte to differentiate were tested. Differentiation into islet like cell was induced by a 15-day protocol with or without ECM gel. Pancreatic characteristics were evaluated with immunofluorescence, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry. Insulin content and release in response to glucose stimulation were detected with chemiluminescent immunoassay system.

RESULTSSixteen MSCs were isolated from 42 term human UCB units (38%). Human UCB-MSCs expressed MSCs specific markers and could be induced in vitro into osteoblast and adipocyte. Islet like cell clusters appeared about 9 days after pancreatic differentiation in the inducing system with ECM gel. The insulin positive cells accounted for (25.2 +/- 3.4)% of the induced cells. The induced cells expressed islet related genes and hormones, but were not very responsive to glucose challenge. When MSCs were induced without ECM gel, clusters formation and secretion of functional islet proteins could not be observed.

CONCLUSIONSHuman UCB-MSCs can differentiate into islet like cells in vitro and ECM gel plays an important role in pancreatic endocrine cell maturation and formation of three dimensional structures.

C-Peptide ; analysis ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Extracellular Matrix ; physiology ; Fetal Blood ; cytology ; Flow Cytometry ; Fluorescent Antibody Technique ; Glucagon ; analysis ; Humans ; Insulin ; analysis ; secretion ; Insulin-Secreting Cells ; cytology ; Mesenchymal Stromal Cells ; cytology ; Reverse Transcriptase Polymerase Chain Reaction

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; enzymology ; pathology ; virology ; Hepacivirus ; genetics ; Humans ; Liver Neoplasms ; enzymology ; pathology ; virology ; Telomerase ; metabolism ; Viral Core Proteins ; genetics ; metabolism

OBJECTIVETo evaluate the role of adiponectin in regulating tumor necrosis factor alpha (TNF alpha) production and preventing fulminant autoimmunological damage of hepatocytes following concanavalin A (Con A) injection into mice.

METHODSThree days after recombinant plasmids pAA-neo-mAd were injected into the mice via the tail veins, Con A was injected into the mice. Mice transfected with empty pAA-neo vector served as controls. The serum levels of alanine aminotransferase (ALT), TNF alpha and adiponectin were detected, and histological examination of livers was carried out at different time points after the Con A injection. All results were subjected to statistical analyses.

RESULTSHistological examinations showed that the damage in livers of mice with high serum adiponectin levels was milder than that of the controls. The serum levels of ALT and TNF alpha were both lower than those of the controls (P less than 0.01, respectively). Statistical analyses showed the serum levels of ALT was negatively related to the levels of adiponectin in the sera (r=-0.5034).

CONCLUSIONAdiponectin is effective in protecting hepatocytes from Con A-induced immunological injury. The mechanism of this protective effect may be caused by inhibiting the synthesis and/or release of TNF alpha.

Adiponectin ; blood ; pharmacology ; Alanine Transaminase ; blood ; Animals ; Concanavalin A ; adverse effects ; Female ; Immune System Diseases ; chemically induced ; pathology ; prevention & control ; Liver ; drug effects ; pathology ; Liver Diseases ; pathology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; blood

Activation of N-methyl-d-aspartic acid (NMDA) receptor plays an important role in neuronal apoptosis induced by cerebral ischemia but the underlying mechanisms are still unclear. The present study examined the neuroprotection of three chloride blockers in an in vitro cell model of cerebral ischemia established by treatment of cultured rat hippocampal neurons with NMDA. Hoechst 33258 staining and MTT assay were used to detect neuronal apoptosis and cell viability, respectively. The neuroprotective effects of chloride channel blockers on the cell viability and neuronal apoptosis were only observed when the blockers were applied before NMDA exposure. In comparison with DIDS, SITS showed more potent protective effect in a dose-dependent manner, whereas NPPB showed no significant neuroprotective effect. The results demonstrate that pretreatment with both SITS and DIDS have protective effect against neuronal apoptosis, which is achieved by blocking both NMDA receptor and chloride channel.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid ; pharmacology ; 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid ; pharmacology ; Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Bisbenzimidazole ; chemistry ; Cell Survival ; drug effects ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; Hippocampus ; cytology ; Microscopy, Fluorescence ; N-Methylaspartate ; pharmacology ; Neurons ; chemistry ; cytology ; drug effects ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

BACKGROUNDHuman umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) could be induced to differentiate into insulin producing cells (IPCs) in vitro, which have good application potential in the cell replacement treatment of type-1 diabetes. However, the mechanisms regulating this differentiation have remained largely unknown. Notch signaling is critical in cell differentiation. This study investigated whether Notch signaling could regulate the IPCs differentiation of human UCB-MSCs.

METHODSUsing an interfering Notch signaling protocol in vitro, we studied the role of Notch signaling in differentiation of human UCB-MSCs into IPCs. In a control group the induction took place without interfering Notch signaling.

RESULTSHuman UCB-MSCs expressed the genes of Notch receptors (Notch 1 and Notch 2) and ligands (Jagged 1 and Deltalike 1). Human UCB-MSCs with over-expressing Notch signaling in differentiation resulted in the down-regulation of insulin gene level, proinsulin protein expression, and insulin-positive cells percentage compared with the control group. These results showed that over-expressing Notch signaling inhibited IPCs differentiation. Conversely, when Notch signaling was attenuated by receptor inhibitor, the induced cells increased on average by 3.06-fold (n = 4, P < 0.001) in insulin gene level, 2.60-fold (n = 3, P < 0.02) in proinsulin protein expression, and 1.62-fold (n = 6, P < 0.001) in the rate of IPCs compared with the control group. Notch signaling inhibition significantly promoted IPCs differentiation with about 40% of human UCB-MSCs that converted to IPCs, but these IPCs were not responsive to glucose challenge very well both in vitro and in vivo. Hence, further research has to be carried out in the future.

CONCLUSIONSNotch signaling may be an important mechanism regulating IPCs differentiation of human UCB-MSCs in vitro and Notch signaling inhibition may be an efficient way to increase the number of IPCs, which may resolve the shortage of islet of cell replacement treatment of type-1 diabetes.

Animals ; Cell Differentiation ; Fetal Blood ; cytology ; Humans ; Insulin ; biosynthesis ; Male ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Receptors, Notch ; physiology ; Signal Transduction ; physiology

Gentiana is an important but complicated group in Gentianaceae. The genus covers numerous medicinal plants which are difficult to be identified. In the present study, several medicinal species in Gentiana from Yunnan province, including G. rigescens, G.rhodantha, and G. delavayi, were sequenced using the Illumina HiSeq 2500 system. Three complete chloroplast genome sequences were obtained after assembly and annotation. According to several published genome sequences of G. crassicaulis, the DNA super-barcoding of species in Gentiana was preliminarily carried out. The results revealed that chloroplast genomes of the three species were conservative with short lengths(146 944, 148 992, and 148 796 bp, respectively). The genomes encoded 114 genes, including 78 protein-coding genes, 30 tRNA genes, 4 rRNA genes, and 2 pseudogenes. Furthermore, these medicinal species in Yunnan province were identified using DNA super-barcoding based on chloroplast genomes. The results showed that the Gentiana species could be gathered into monophyletic branches with a high support value(100%). It indicated that DNA super-barcoding possessed obvious advantages in discriminating species in complicated genera. This study is expected to provide a scientific basis for the identification, utilization, and conservation of Gentiana species.
China ; DNA ; Genome, Chloroplast/genetics* ; Gentiana/genetics* ; Phylogeny

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Hemorrhagic Fevers, Viral ; epidemiology ; Hospitalization ; Humans ; Male ; Middle Aged

OBJECTIVETo compare the efficacy of all-trans retinoic acid (ATRA) combining chemotherapy and As4S4 with ATRA combining chemotherapy for the maintenance treatment of patients with acute promyelocytic leukemia (APL).

METHODSSixty patients with APL induced to complete remission by ATRA and consolidated by chemotherapy were randomly divided into two groups. Thirty patients as As4S4 group received ATRA + As4S4 + chemotherapy, and another thirty patients as non-As4S4 group were treated only with ATRA + chemotherapy as maintenance therapy. The therapeutic effects, side effects and PML-RARalpha gene expression were analyzed.

RESULTSThe three-year continuous complete remission (CCR) rate was 90.0% for As4S4 group and 61.1% for non-As4S4 group, the difference being statistically significant. Significant difference was also found in the positive rate of PML-RARalpha fusion gene between the two groups. The side effects were mild.

CONCLUSIONAPL patients in maintenance therapy with ATRA + 6-MP + MTX + As4S4 can obtain a higher CCR.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; adverse effects ; Arsenicals ; therapeutic use ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; Male ; Middle Aged ; Remission Induction ; Sulfides ; therapeutic use ; Treatment Outcome ; Tretinoin ; therapeutic use

OBJECTIVETo investigate the effects of the different treatments of deep venous thrombosis (DVT) of lower extremities on the incidence of the pulmonary embolism (PE).

METHODS201 patients (97 males and 104 females, mean age 60.4 years ranged from 24 to 83) from August 2002 to June 2008 with DVT were retrospectively reviewed and divided into 3 groups based on different treatment, including anticoagulants plus thrombolytics alone (group 1), thrombectomy plus anticoagulants plus thrombolytics (group 2) and anticoagulants plus thrombolytics after delivery of inferior vena cava (IVC) filter (group 3) respectively. One hundred and seventy-four cases had left lower limb DVT, 24 cases had right lower limb DVT and 3 cases had both sides of lower limb DVT. Different incidence of PE in different period (7-14 d in hospital and follow-up after discharge) were calculated. Effects of the three different treatment methods of DVT on the incidence of PE were studied.

RESULTSFor in-patients, the prevalence of symptomatic PE was 2.8% (3/107) in the group of receiving anticoagulants plus thrombolytics alone, but in the other two groups, no symptomatic PE happened. There was no significant difference in incidence of symptomatic PE among the 3 groups (P=0.425). For patients discharged, after 6 to 72-month follow-up (mean 24-month), we found that no PE happened in group 1 and group 2, while in group 3, the incidence of PE was 2.4% (1/42). There was also no significant difference (P=0.656) among 3 groups.

CONCLUSIONSThere is no significant difference in relation to the incidence of PE in these 3 groups. Therefore vena cava filter implantation should be restricted to optimal indication.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Incidence ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Pulmonary Embolism ; etiology ; prevention & control ; Retrospective Studies ; Venous Thrombosis ; complications ; therapy