Blood Vessels ; anatomy & histology ; Facial Nerve ; blood supply ; Humans

OBJECTIVETo investigate the regulatory mechanism of the transcription of tumor metastasis suppressor gene TMSG-1.

METHODSLuciferase reporter assay and site-directed mutagenesis were used to analyze the regulatory region of TMSG-1. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were carried out to verify the interaction of KLF6 and Sp1 with the regulatory region of TMSG-1. Co-immunoprecipitation (CoIP) was performed to analyze the interaction between KLF6 and Sp1. TMSG-1 and wt-KLF6 mRNA expressions in cells with different metastatic capacities were quantitated by real-time PCR. Cell invasive capability was determined by Matrigel invasion assay.

RESULTSA 63 bp inducible regulatory region (+59 bp - +123 bp) in exon 1 was identified by luciferase assay using reporter plasmids with a series of TMSG-1 regulatory region deletions. Mutations in KLF6/Sp1 binding sites of this region resulted in a decrease of luciferase activity, while cotransfection with KLF6 or Sp1 expressing plasmids led to a remarkable increase of luciferase activity. EMSA and ChIP demonstrated that KLF6 as well as Sp1 interacted with this region. CoIP also indicated a possible interaction between KLF6 and Sp1 proteins. In the highly metastatic cell sublines, a low level of wild type KLF6 was associated synchronously with a low TMSG-1 level. Prostate carcinoma cells overexpressing KLF6 exhibited a higher TMSG-1 level and a lower invasive capability.

CONCLUSIONSTranscription factor complex of KLF6 and Sp1 may participate in the inducible transcriptional regulation of TMSG-1, and a decreased wild type KLF6 expression is likely associated with a low TMSG-1 level in the highly metastatic cell sublines.

Binding Sites ; genetics ; Cell Line, Tumor ; Electrophoretic Mobility Shift Assay ; Humans ; Immunoprecipitation ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mutagenesis, Site-Directed ; Mutation ; Neoplasm Invasiveness ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Sp1 Transcription Factor ; genetics ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transcriptional Activation ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo observe the effects of mesenteric lymph duct (MLD) ligation on erythrocyte rheology in acute hemorrhagic rats.

METHODSTwenty male Wistar rats were randomly divided into hemorrhage group and ligation group (n = 10). Blood (one fourth of body whole blood volume) was withdrawn through right common carotid arteries after rats were anesthetized. In ligation group, the MLD was ligated after hemorrhage, and only threading under the MLD in hemorrhage group. The survival situation at 24 h was recorded. After 24 h, survival rats were anesthetized again, blood sample was withdrawn through left common carotid artery rapidly. And the erythrocyte sedimentation rate (ESR), electrophoresis of erythrocytes, hematocrit (Hct) were determined in blood samples of before and after hemorrhage, the erythrocytes aggregation and deformability indices were calculated.

RESULTSIt showed that the ligation group survival (9 rats alive) was slightly better than that in hemorrhage group (6 rats alive). The results of erythrocyte rheology indices showed that the ESR, K value of equation, K value of emendation and electrophoresis time in hemorrhage group and ligation group were higher or longer than those before hemorrhage, the erythrocyte deformability was reduced significantly, respectively. And the erythrocytes aggregation index in hemorrhage group was increased, the electrophoresis length and migration of erythrocyte in hemorrhage group were lower than those before hemorrhage, respectively. But compared with hemorrhage group, the ESR, K value of equation, K value of emendation, erythrocytes aggregation index and electrophoresis time in ligation group were lower, the electrophoresis lenght, migration and deformability of erythrocyte were increased significantly.

CONCLUSIONThe results indicate that the higher erythrocyte aggregation ability, lower electrophoresis function and deformability are caused by acute hemorrhage in rats, and the MLD ligation can improve the abnormal erythrocyte rheology.

Animals ; Disease Models, Animal ; Erythrocyte Deformability ; Erythrocytes ; pathology ; Hemorrhage ; surgery ; Ligation ; Lymphatic Vessels ; surgery ; Male ; Mesentery ; surgery ; Rats ; Rats, Wistar ; Rheology ; Shock, Hemorrhagic ; surgery

OBJECTIVETo observe the effects of mesenteric lymph drainage on erythrocyte rheology and blood viscosity in hemorrhagic shock rats.

METHODSWistar rats were randomly divided into sham-shock group, shock group (establishing hemorrhagic shock model), drainage group (establishing hemorrhagic shock model plus drainaging shock mesenteric lymph from hypotension 1 h). At 3 h of hypotension or corresponding time, blood samples were harvested from the abdominal aorta for determining the erythrocytic parameters, erythrocyte electrophoresis, erythrocyte sedimentation rate (ESR) and blood viscosity, and the erythrocytes aggregation index and erythrocyte deformability index were calculated.

RESULTSCompared with the sham-shock group, the red cell contents, hematocrit (HCT), hemoglobin (Hb), mean corpuscular hemoglobin concentration (MCHC), erythrocyte electrophoretic rate and mobility, erythrocyte deformability index, whole blood viscosity, whole blood relative or reduced viscosity at low and high shear rates in shock group were observably lower, and mean corpuscular volume, electrophoretic time of erythrocyte, ESR, K value of equation and K value of emendation, erythrocytes aggregation index, plasma viscosity in shock group were increased markedly; the MCHC, erythrocyte electrophoretic rate and mobility, whole blood viscosity, whole blood relative viscosity at low and high shear rates in drainage group were reduced, and the red blood cell volume distribution width -SD (RDW-SD) was increased remarkably. At the same time, in drainage group, the HCT, RDW-SD, erythrocyte deformability index, whole blood viscosity and relative viscosity at low and high shear rates were higher, the ESR, K value of equation and K value of emendation, erythrocytes aggregation index, plasma viscosity were lower than that of shock group.

CONCLUSIONThe results indicate that the mesenteric lymph drainage could improve the erythrocyte rheological behavior, as a result, improve the hemorrheological properties in hemorrhagic shock rats.

Animals ; Blood Viscosity ; Drainage ; methods ; Erythrocyte Aggregation ; Erythrocyte Deformability ; Lymph ; Male ; Mesentery ; Rats ; Rats, Wistar ; Rheology ; Shock, Hemorrhagic ; blood ; therapy

Objective To investigate the expressions of S100B and calcitonin gene related peptide (CGRP) and the pathologic alterations of the hippocampus in kainic acid (KA)-induced epileptic rats. Methods Male SD rats were randomly divided into control group (n=8) and model group (n=40).Animal models of temporal lobe epilepsy were established by intracerebroventricular injection of KA; the same volume of saline was injected into the rats in the control group. Hippocampal tissues within various phases after seizures (6, 12, 24 and 72 h, and 24 h after the success of model making) were performed Nissl staining, Timm staining and immunohistochemical staining. The expressions of S100B and CGRP were observed, and the pathologic alterations of the hippocampal neurons and glial cells were studied.Results All rat models were successfully induced with epileptic seizures. Nissl staining showed that pyknotic neuronal necrosis appeared in the CA3 area of the hippocampus in the model group with cell body atrophy and disappearance of Nissl bodies 1 week after the injection. Timm staining showed that brown particles showed stripped distribution in the CA3 area of the hippocampus and some brown particles in the molecular layer of fascia dentate. Immunohistochemical staining indicated that significant neurons lost and gliosis appeared after seizures with abundant expressions of S100B and CGRP.Conclusion KA-induced epileptic rats express abundant S100B and CGRP and appear such pathological changes as disappearance of Nissl bodies and mossy fiber sprouting, indicating that both S100B and CGRP participate in the onset of epilepsy.

OBJECTIVETo study the activity, protein and gene expression of renal HK-ATPase (HKA) in rats subchronic exposed to trimethyltin chloride (TMT).

METHODSIn subchronic toxic test (14-week), 55 female SD rats (age, 6 weeks) were divided randomly into 5 groups: control, low, medium, high and super high dosage, respectively, which drank water with TMT of 0, 8.20, 32.81, 131.25 and 262.50 microg x kg(-1) x d(-1) for 14 weeks. Then serum K+ levels were measured; the activities of HK-ATPase (HKA) in kidneys were detected by the method of determinated phosphorus content; Western Blot assay and real-time PCR were used to exam the protein and mRNA expression levels of HKA in kidneys, respectively.

RESULTSThe serum K+ level in super-high dosage group was (5.6 +/- 0.4) mmol/L, which was significantly lower than that [(6.9 +/- 0.3) mmol/L] in control group (P < 0.01). The HKA enzymatic activity of kidneys in low and super high dosage groups was 4.50 +/- 1.45 and 4.55 +/- 0.72 micromolPi x mg prot(-1)h(-1), respectively, which were significantly lower than that (6.55 +/- 0.77 micromol Pi x mg prot(-1) h(-1)) in control group (P < 0.05).

CONCLUSIONWhen rats were exposed subchronic to TMT, the renal HKA activity could reduce, but the expression levels of HKA protein and mRNA did not decrease.

Animals ; Female ; Gene Expression ; H(+)-K(+)-Exchanging ATPase ; genetics ; metabolism ; Kidney ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Toxicity Tests, Subchronic ; Trimethyltin Compounds ; toxicity

OBJECTIVETo report an X-linked dominant Charcot-Marie-Tooth disease (CMTX) Chinese family with vocal cord paresis and to identify the mutation of gap junction protein beta 1 gene (GJB1).

METHODSPart of the family members with dysphagia, dysphonia and lethal respiratory failure were studied through flexible laryngoscope, clinical, brain MRI and electrophysiological examinations. After excluding large fragment tandem duplication containing peripheral myelin protein 22 gene (PMP22), direct sequencing was performed to analyze the mutation of the GJB1 gene in 5 patients including the proband, 5 unaffected family members and 50 unrelated healthy individuals.

RESULTSEight members spanning 3 generations in this family were affected with CMTX characterized by progressive atrophy and weakness of the anterior tibial and peroneal muscles, especially in the proband. Vocal cord paresis was observed through flexible laryngoscope in total of 4 affected members with dysarthria and dysphagia, 2 of them died of severe respiratory failure due to complete bilateral vocal cord involvement. Normal brain MRI was observed in the proband. The electrophysiological data showed predominant demyelization involving the motor and sensory nerves in the proband. DNA sequencing revealed a de novo c.186 C>G missense mutation in exon 2 of the GJB1 gene, the mutation cosegregated with phenotype.

CONCLUSIONRespiratory failure associated with vocal cord involvement may be a rare and severe symptom in CMTX. The present report provides further evidence for clinical and genetic heterogeneity in the X-linked Charcot-Marie-Tooth disease.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Charcot-Marie-Tooth Disease ; genetics ; Connexins ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; Myelin Proteins ; genetics ; Pedigree ; Vocal Cord Paralysis ; genetics ; Young Adult

OBJECTIVETo observe the change of erythrocyte theology in rabbits with acute renal failure (ARF).

METHODSThirty-eight healthy rabbits were randomly divided into control group (n = 8), model group (establishing ARF model via intramuscular injection 1% HgCl2, and divided into 12 h, 24 h, 48 h subgroups, all n = 10), the arterial blood sample was taken out through carotid artery at corresponding times after anesthetization with urethane, for detecting the indices of renal function and erythrocyte rheology.

RESULTSThe levels of urea and creatinine in plasma of model rabbits at 12 h, 24 h and 48 h were higher than those of control group, and there was a rise trend along with the time extension. The erythrocytes electrophoresis time at 12 h of model group was higher, the electrophoresis rate and migration rate of erythrocytes were lower compared with those of control group; the erythrocytes electrophoresis time at 24 h of model group was lower and the electrophoresis rate and migration rate were higher compared with those of model group at 12 h; and there were no statistical differences in erythrocytes electrophoresis indices between model group at 48 h and other groups. Meanwhile, there was a rise trend in erythrocyte sedimentation rate (ESR), K value of equation and emendation along with the time extension of ARF, but these indices only at 48 h of model group was lower significantly than that of control group. There were no statistical differences in aggregation index and deformability index of erythrocytes among groups.

CONCLUSIONDuring the process of ARF, the erythrocytes electrophoresis time lengthen and electrophoresis rate and migration rate decrease at early stage, and these indices gradually return to normal; the indices of ESR increase gradually.

Acute Kidney Injury ; blood ; physiopathology ; Animals ; Erythrocyte Indices ; Erythrocytes ; physiology ; Hemorheology ; Rabbits

OBJECTIVETo observe the changes of blood viscosity and erythrocyte rheology in mice after acute hypoxic hypoxia (AHH).

METHODSThirty-two Kui-ming mice were randomly divided into control group, AHH group (duplicating AHH model, and divided into 5 min, 8 min, 11 min subgroups), the blood sample was taken out from heart after neck dislocation at corresponding times, for detecting the blood viscosity and erythrocyte rheology indices.

RESULTSCompared with control group, the whole blood viscosity at different shears, whole blood reduced viscosity, whole blood relative viscosity were lower and the erythrocytes aggregation index was higher in AHH 5 min group; the electrophoresis time was longer and the electrophoresis length, migration of erythrocyte were lower in AHH 8 min and AHH 11 min groups. The whole blood reduced viscosity, whole blood relative viscosity, erythrocytes aggregation index in AHH 8 min group were higher, and the erythrocyte deformability index was lower significantly than that of AHH 5 min group, respectively.

CONCLUSIONThese data suggested that the AHH could induce the blood viscosity and electrophoresis ability.

Animals ; Blood Viscosity ; Erythrocyte Aggregation ; Erythrocyte Deformability ; Erythrocyte Indices ; Erythrocytes ; physiology ; Hypoxia ; blood ; etiology ; Male ; Mice ; Mice, Inbred Strains

OBJECTIVETo investigate the pathophysiological role of CXCL16 in immunological liver injury induced by Bacille de Calmette et Guerin (BCG) and lipopolysaccharides (LPS).

METHODSImmunological liver injury was induced by BCG and LPS in mice, and the expression of CXCL16 was detected in the liver tissues by real-time quantitative PCR and immunohistochemical examination. The relationship of the expression of CXCL16 and the extent of hepatic necrosis was investigated histopathologically and immunohistochemically. Mononuclear cells were isolated from the liver tissues and their numbers were counted; T lymphocytes populations in the liver tissue were also analyzed with FACS.

RESULTSThe immunological liver injury model was successfully created. Up-regulation of CXCL16 in injured livers correlated with the extent of liver injury and the amountmononuclear cell infiltrations.

CONCLUSIONThese findings suggest that up-regulation of CXCL16 was closely correlated with liver injury extent during the immunological liver injury induced by BCG-LPS in mice, and intrahepatic recruitment of specific lymphocytes might be an important mechanism of liver injury.

Animals ; Chemical and Drug Induced Liver Injury ; Chemokine CXCL16 ; Chemokine CXCL6 ; Chemokines, CXC ; biosynthesis ; genetics ; Lipopolysaccharides ; Liver Diseases ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Mycobacterium bovis ; Receptors, Scavenger ; biosynthesis ; genetics