OBJECTIVE: To investigate the effect of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, on the activation of hepatic stellate cells (HSCs) and its mechanism. METHODS: Primary HSCs isolated from SD rats were cultured and treated with different concentrations (1, 3 or 10micromol/L) of ADMA for various periods (12 approximately 48h). Expression of alpha-smooth muscle actin (alpha-SMA) and synthesis of type-I collagens in HSC were determined. Messenger RNA levels of the transforming growth factor-beta1 (TGF-beta(1)) in the HSCs were determined using RT-PCR. Intracellular reactive oxidant species (ROS) production was measured using oxidant-sensitive fluorescent indicator. Activation of nuclear factor-kappaB (NF-kappaB) was detected by electrophoretic mobility shift assay (EMSA). RESULTS: ADMA could increase alpha-SMA-positive cells ratio and Type I collagens production of HSCs in a concentration- and time-dependent manner, concomitant with the increase of the TGF-beta(1) mRNA level. Treatment with ADMA (10micromol/L) significantly increased the intracellular ROS production and activated NF-kappaB. Such effects of ADMA on the level of TGF-beta(1) mRNA could be markedly attenuated by pretreatment with antioxidant pyrrolidine dithiocarbamate (25micromol/L). CONCLUSION ADMA can induce the HSC activation by increasing TGF-beta(1) expression through ROS-NF-kappaB-dependent pathway. Therefore, ADMA should be a novel and endogenous activator of HSC, which may be involved in the development of liver fibrosis.
Actins ; biosynthesis ; Animals ; Arginine ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Collagen Type I ; metabolism ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Hepatocytes ; cytology ; drug effects ; metabolism ; Male ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics

Objective: To investigate the effect of rhubarb on gastrointestinal blood perfusion in critical illness and hemorrhagic shocked rats.Methods: Clinical Study: Sixty-four septic patients, who suffered from stress ulcer, were treated with rhubarb at a dose of 25 mg/kg. Twenty-five non-septic patients were taken as control. The gastrointestinal perfusion was evaluated by intramural pH (pHi). Animal study: SD rats were anesthetized with intraperitoneal sodium pentobarbital at a dose of 20 mg/kg. Blood-letting were performed in the animals. Blood pressure reduced to 5.32 kPa and maintained for 120 mins. They were resuscitated at the end of shock by reinfusing all of the shed blood. The rats were randomly divided into four groups: Normal control, shock group, therapeutic group (shocked rats were treated with 50 mg/kg rhubarb at the end of shock) and rhubarb group (normal rats were treated with rhubarb). Laser Doppler was applied to estimate the gastrointestinal blood perfusion. Results: Clinical Study: The gastrointestinal pHi in septic patients was much lower than that in the control, whereas rhubarb could obviously elevate gastrointestinal pHi (P<0.001). In addition, rhubarb also had good effect on gastric hemorrhage caused by stress ulcer. Animal Study: Although the shocked rats were resuscitated completely, their gastrointestinal blood perfusion was much lower than that in the control. Rhubarb could significantly improve the blood perfusion in gastrointestinal mucosa and mesentery (P<0.01). Furthermore, rhubarb also increase the gastrointestinal perfusion in normal rats. Conclusion: Rhubarb could improve gastrointestinal blood perfusion in critical illness and shocked rats.

BACKGROUNDPreS/S gene was derived from hepatitis B virus (HBV) integration fragment of human hepatocellular carcinoma genome, containing the promoter of preS/S and the C terminal truncated preS/S open reading frame. PreS/S protein may have important roles in processing hepatoma in some HBV-infected patients. The aim of the study was to study the activity of HBV preS/S protein on proliferating cell nuclear antigen (PCNA) promoter and to localize compartment of the preS/S protein in the liver cell line L02.

METHODSThe authors studied the effect of the 3 -truncated preS/S on human PCNA promoter by co-transfecting the expression plasmids of luciferase reporter gene, used the immunohistochemical method to localize the preS/S protein.

RESULTSThe expression product of the plasmid, pKSH7C-HpaI which contained the 3 -truncated preS/S and the flanking cellular sequences, stimulated the expression of PCNA promoter dose dependently,and its effect was 0.5 folds higher than control. Immunohistochemistry showed that the preS/S protein located in the cytosolic region of the liver cell line L02.

CONCLUSIONSThe HBV preS/S protein could stimulate the PCNA promoter of the liver cell, its effect was not direct, which suggests that the effect of preS/S protein on PCNA promoter was probably through the cell signal transduction pathway.

Carcinoma, Hepatocellular ; genetics ; virology ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; virology ; Proliferating Cell Nuclear Antigen ; genetics ; Promoter Regions, Genetic ; Protein Precursors ; genetics ; Tumor Cells, Cultured ; Virus Integration

The complex field cure instrument is a new medical instrument with which an experiment was carried out. Rats were continuously irradiated by the complex field for 90 days, with a day's total dose of 285.9 M.T.G. while other rats weren't irradiated for control group. The animals were respectively killed at 7d, 14d, 30d, 60d and 90d, and their blood samples were taken for cell and humoral immune analysis. The results show that values of lymphocyte transform rate, soluble receptor (SIL-2R), total hemolytic complement levels (CH50) and immunoglobulin (A.G.) after irradiation are more than those of the control group having proved that the instrument may improve immune function of rats.
Animals ; Female ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulins ; blood ; Lymphocyte Activation ; Lymphocyte Count ; Male ; Physical Therapy Modalities ; instrumentation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin-2 ; blood ; Time Factors

OBJECTIVE: To explore the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus after ovariectomy in mice. METHODS: BDNF levels were detected by immunohistochemistry combined image analysis in hippocampal CA regions and dentate gyrus of ovariectomized mice. RESULTS: The expression of BDNF in hippocampus of mice decreased significantly after the ovariectomy after 4 days. The recovery BDNF expression started 14 days after the ovariectomy and after 28 days, the expression of BDNF in hippocampus recovered to the normal level. CONCLUSION The decrease of estrogen in ovariectomized mice can weaken the expression of BDNF in hippocampus during the early stage.
Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Dentate Gyrus ; metabolism ; Estrogens ; blood ; Female ; Hippocampus ; metabolism ; Mice ; Ovariectomy ; RNA, Messenger ; biosynthesis ; genetics

BACKGROUNDParoxysmal kinesigenic dyskinesia (PKD) is characterized by recurrent brief episodes of chorea and dystonia induced by sudden movement. Whether the central nervous system is hyper- or hypoexcitable in PKD remains undetermined. The aim of our study was to compare the somatosensory evoked potential (SEP) recovery cycle, a marker of somatosensory system excitability, in PKD patients and controls.

METHODSTwenty-four PKD patients (mean age of (20.0±5.3) years; 21 males, 3 females) and 18 control age-matched subjects (mean age of (22.0±5.0) years; 17 males, 1 female) were studied. The stimuli were delivered to the median nerve in the affected dominant arm in patients and in the dominant arm in controls. The change in SEP amplitude was measured after paired electrical stimulation at interstimulus intervals (ISIs) of 5, 20, and 40 ms. The SEPs evoked by S2 (test stimulus) were calculated by subtracting the response to S1 (the conditioning stimulus) from the response to a pair of stimuli (S1+S2), and their amplitudes were compared with those of the control response (S1) at each ISI. Analysis of variance (ANOVA) or equivalent was used for non-parametric data.

RESULTSIn patients, the P27 amplitude after the single stimulus (S1) was significantly larger than that after the control stimulus. The (S2/S1)×100 ratio for P14 and N30 SEPs did not differ significantly between PKD patients and normal subjects at ISI of 5 ms but were significantly higher in patients at ISIs of 20 and 40 ms (P<0.05).

CONCLUSIONSSomatosensory system disinhibition takes place in PKD. The finding of reduced suppression of different SEPs, each thought to have a different origin, suggests an abnormality of intracortical and subcortical inhibitory circuits.

Adolescent ; Adult ; Case-Control Studies ; Dystonia ; physiopathology ; Evoked Potentials, Somatosensory ; physiology ; Female ; Humans ; Male ; Young Adult

OBJECTIVETo investigate the efficacy of imatinib in the treatment of chronic myeloid leukemia (CML) and analyse the treatment outcome and related factors.

METHODSNinety five CML patients were treated with imatinib in our hospital from May 2002 to May 2006. The outcomes and related factors were analysed.

RESULTS(1) One year after therapy, there were 95.5% of chronic phase (CP) patients achieved complete hematologic response (CHR). Fifty-two patients with complete cytogenetic dates were divided into primary-therapy group (n = 19) and secondary-therapy group (n = 33). The major cytogenetic responses (MCyR) at 6-, 12-, 18-, 24- and 30-months after therapy for the former group were 84.2%, 84.2%, 89.5%, 89.5% and 94.7%, and for the latter group were 36.4%, 39.4%, 39.4%, 39.4% and 39.4%, respectively (P < 0.01). The expected survival at 12-, 24-, 36- and 50-month after imatinib treatment for CP group was (98.1 +/-1.9)%, (87.8 +/- 7.1)%, (81.9 +/- 8.7)% and (81.9 +/- 8.7)%, respectively. (2) Twelve month after therapy, there are 70% of accelerated phase (AP) patients achieve CHR and 10% get MCyR. The expected survival at 12-, 24- and 36-month after imatinib treatment for AP group was (63.0 +/- 17.7)%, (15.8 +/- 14.3)% and (15.8 +/- 14.3)%, respectively. (3) Six month after therapy, 57.9% of blast crisis (BC) patients achieve CHR, with the expected survival at 12- and 24-month of (40.6 +/- 12.3)% and 0, respectively. (4) COX analysis CP group indicated that imatinib therapy administered for previously untreated was an independent favorable prognostic factor. Conclusion (1) Imatinib as a primary treatment for CP CML can significantly improve the survival time as compared with that AP or BC patients or with that used in previously treated patients. (2) Imatinib could induce hematologic, even cytogenetic response to a certain extent, in CP or BC patients and prolong the survival time.

Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Male ; Middle Aged ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo illustrate the early alteration of plasminogen activator inhibitor-1 (PAI-1) in the recipients of hematopoietic stem cell transplantation (HSCT) and explore its clinical significance in transplantation-associated thrombotic complications.

METHODSNinety-five patients undergoing HSCT were enrolled in this study. PAI-1 level and other hemostatic parameters were measured by enzyme linked immunosorbent assay (ELISA) in platelet poor plasma samples from patients on conditioning therapy and then weekly until four weeks after HSCT.

RESULTSSignificant increase in PAI-1 was detected after conditioning treatment, followed by a diminution in the very week on transplantation (week 0), then increased with in time after transplantation. According to the occurrence of transplant-associated complications, patients were classified into four groups: thrombus group \[veno-occlusive disease (VOD) (n = 5), thrombotic microangiopathy (TMA) n = 1\], aGVHD group (n = 29), infection group (n = 19) and non-complication group (n = 41). One of 30 patients (3.3%) was diagnosed as thrombus in the auto-HSCT group, while five of 65 patients (7.7%) did in the allo-HSCT group. PAI-1 level of thrombotic patients was significantly increased compared with non-thrombotic subjects, and the patients without thrombotic complications have higher PAI-1 level in the allo-HSCT group than in auto-HSCT group. All the patients with complications presented with significantly increased PAI-1 compared with those with no complications (P < 0.05). The six patients with thrombotic complications showed extremely elevated PAI-1 \[(62.8 +/- 7.5) microg/L\] compared with that of aGVHD patients \[(45.1 +/- 9.1) microg/L\] or infection patients \[(50.0 +/- 11.2) microg/L\] post-HSCT (P < 0.05).

CONCLUSIONThe increase in plasma PAI-1 may be a specific mark for transplantation-associated thrombotic complications. Increased PAI-1 reflects the development of thrombotic complications. Extreme elevation of PAI-1 contributes to the early diagonsis of VOD and TMA after HSCT.

Hematopoietic Stem Cell Transplantation ; Hemostasis ; Humans ; Thrombosis ; Thrombotic Microangiopathies

OBJECTIVETo observe the effect of ligand of peroxisome proliferators-activated receptor gamma (PPAR gamma) 15d-PGJ2 on the proliferation and activation of hepatic stellate cells (HSC) and to study the role played by PPAR gamma during the process of HSC activation.

METHODSBy using RT-PCR and cell culture, we investigated the effects of 5 micro mol/L and 10 micro mol/L 15d-PGJ2 on culture-activated HSC and on PDGF-induced HSC proliferation, production of extracellular matrix and expression of chemokines.

RESULTSThe expression of alpha-SMA was significantly suppressed by 5mumol/L 15d-PGJ2, and the expression of PPAR gamma was significantly higher in the 15d-PGJ2 treated group than in the untreated group (0.64+/-0.03 vs 0.09+/-0.01, t=36.0517, P<0.01); PDGF-induced HSC proliferation was dose-dependently suppressed by 15d-PGJ2; the expressions of PPAR gamma in 5 micro mol/L and also in 10 micro mol/L 15d-PGJ2 plus PDGF pre-treated group increased much more than those in the PDGF-treated group (0.03+/-0.02 vs 0.60+/-0.03, t=42.6616, P<0.01 and 0.03+/-0.02 vs 0.69+/-0.04, t=33.83, P<0.01); the expressions of alpha-SMA, alpha 1 (I)-collagen and MCP-1 were suppressed.

CONCLUSIONActivation of PPAR gamma can modulate pro-fibrotic and pro-inflammatory roles of HSC and the increased expression of PPAR gamma may become a new target for antifibrosis.

Animals ; Cell Differentiation ; Cell Proliferation ; drug effects ; Cells, Cultured ; Hepatic Stellate Cells ; cytology ; metabolism ; Male ; PPAR gamma ; metabolism ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; Rats ; Rats, Wistar

To screen factors related to spermatogonial stem cell (SSC) proliferation, and to investigate the mechanism of infertility caused by cryptorchidism, ten-day-old Kunming (KM) mice were used and experimental cryptorchidism was conducted. On the 35th day after cryptorchid operation, the left testes were fixed in Bouin's fluid and used for histological analysis. The testes of 45-day-old mice were subjected to the same histological analysis, and it was found that they contained germ cells at every stage of development, from SSCs to sperm, indicating that the animals were fully sexually mature at this age. While in experimental cryptorchid mice, the spermatogenesis was arrested at the stage of spermatocytes, and only spermatogonia and primary spermatocytes were present in cryptorchid testes. The proportion of spermatogonia to other types of germ cells was much higher than that in sexually mature mice. On the other hand, the right testes were used for proteomic analysis. The total protein in testes was extracted on the 35th day after cryptorchid operation. The differentially expressed proteins in cryptorchid mice and sexually mature mice were screened and compared by the proteomic techniques. Through the separation of two-dimensional gel electrophoresis (2-DE), 20 differential protein spots were found, and 9 of them were digested and identified by the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrum. In cryptorchid mice, 6 out of 9 proteins were down-regulated, and 3 were up-regulated. Among these proteins, 4 proteins were identified, and they were Stathmin, phosphatidylethanolamine-binding protein1 (PEBP1), HES-related basic helix-loop-helix protein (HERP), and one unnamed protein (we temporarily named it Px). More Stathmin, PEBP1 and Px were expressed in sexually mature mice than in experimental cryptorchid mice. But HERP1 was the other way round. In the present study, we have screened 4 proteins related to cryptorchidism. It is helpful to study the mechanism of SSC proliferation and infertility caused by cryptorchidism.
Animals ; Cryptorchidism ; metabolism ; Male ; Membrane Proteins ; analysis ; Mice ; Phosphatidylethanolamine Binding Protein ; analysis ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stathmin ; analysis ; Testis ; chemistry