Aim To study the effect of melatonin on expression and activity of myosin light chain kinase in the artery wall of atherosclerotic rabbits.Methods The rabbit model of atherosclerosis was induced by a high-cholesterol diet.The blood lipid levels were assayed in the serum of each group.MLCK expression was detected by Western blot and immunohistochemical method.MLCK activity was measured by ?-32P-ATP incorporation into myosin light chain.Results The atherosclerosis model was established successfully.The levels of lipids decreased after MLT treatment.After fed with cholesterol for twelve weeks,the expression and activity of MLCK in the artery of atherosclerotic rabbits increased markedly,whereas there was no obvious difference in expression of MLCK in the artery of atherosclerotic rabbits fed with cholesterol and melatonin for twelve weeks compared with that of control.Conclusions It was suggest that high expression and activity of MLCK in the artery might be closely correlated with the development of atherosclerosis.Melatonin played an important role in inhibiting the development of atherosclerosis by decreasing the expression and activity of MLCK.

BACKGROUND/AIMS: Kruppel-like factor 4 (KLF4) is an epithelial-specific transcription factor primarily expressed in the gastrointestinal tract that mediates growth arrest in the colonic epithelium. We tried to find whether KLF4 expression is associated with the progression and differentiation of colorectal cancer. METHODS: We detected KLF4 expression in 109 colorectal specimens (40 normal appearing mucosa, 7 adenomas, and 62 carcinomas) by immunohistochemistry using a tissue microarray. Western blot and RT-PCR analyses were also performed. RESULTS: The upregulation of KLF4 expression in carcinoma tissue was statistically significant (p<0.05) when compared to normal appearing mucosa. The negative and weak positive staining rates in normal appearing mucosa, adenoma, and carcinoma were 42.5%, 71.4%, and 82.3%, respectively, indicating a decreased degree of KLF4 expression over the course of progressive transformation of normal cells into malignant derivatives. KLF4 protein levels showed no correlation with sex, age, or metastatic state (p>0.05), while KLF4 protein expression correlated with the diagnostic stage (p<0.05). Furthermore, strong KLF4 staining was detected in 22.9% (11/48) and 0% (0/14) of well/moderately and poorly differentiated colorectal cancers, respectively. Our results clearly indicate that KLF4 protein expression significantly correlates with the degree of differentiation in colorectal cancers (p<0.05). KLF4 expression in RKO cells is also upregulated by butyrate, an inducer of differentiation. CONCLUSIONS: Downregulation of KLF4 expression may lead to more poorly differentiated tumors.
Adenoma ; Blotting, Western ; Butyrates ; Colon ; Colorectal Neoplasms ; Down-Regulation ; Epithelium ; Gastrointestinal Tract ; Immunohistochemistry ; Kruppel-Like Transcription Factors ; Mucous Membrane ; Transcription Factors ; Up-Regulation

OBJECTIVETo investigate whether there were differences in the clinical characteristics, cytogenetic characteristics, immunophenotype and prognosis in children with B cell type acute lymphoblastic leukemia (B-ALL) carrying different fusion genes.

METHODSThe research included 80 children with B-ALL from Peking University People's Hospital between March 2006 and December 2008. Eighteen children were positive for TEL/AML1, 14 for E2A/PBX1, 11 for BCR/ABL,and 2 cases for MLL/AF4, and 35 cases were negative for all of the 4 fusion genes. Data including clinical characteristics, morphology, immunophenotype and cytogenetic characteristics were collected, and the disease-free survival (DFS) was evaluated. The children were followed up until April 2009.

RESULTSIn the 18 children with TEL/AML1+B-ALL, 66.7% were younger than 5 years old. They had low tumor load. FAB-L2 morphology was commonly observed, but t(12;21) was often absence in these children. Up to now,17 children who survived were disease-free. In the 14 children with E2A/PBX1+B-ALL, the majority were female. Thirteen children showed FAB-L1 morphology. Twelve children showed pre-B-ALL immunophenotype. The EFS was close to 80%. In the 11 children with BCR/ABL+B-ALL, 10 children showed common B type immunophenotype. FAB-L1 and FAB-L2 morphology was found in 4 children respectively. The DFS was less than 20%. Two children with MLL/AF4 positive B-ALL had high tumor load. Their morphologic diagnosis was FAB-L1. Both showed the Pro-B-ALL immunophenotype. One child discontinued treatment at the early stage of chemotherapy, and the other child survived disease-free until now.

CONCLUSIONSThe B-ALL children with different fusion genes have different clinical characteristics, immunophenotypes and prognosis.

Adolescent ; Child ; Child, Preschool ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Gene Fusion ; Homeodomain Proteins ; genetics ; Humans ; Immunophenotyping ; Infant ; Male ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Oncogene Proteins, Fusion ; genetics ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology

OBJECTIVETo investigate the specific antitumor immunity induced by antigen peptide mixture prepared from different T lymphocytic leukaemia cells and the cross-reaction among the mixtures of different cell lines.

METHODSAntigen peptide mixtures were prepared from different leukaemia cell lines and then bound with Hsp70 in vitro. The activation and proliferation of peripheral blood mononuclear cell (PBMC) were observed after the stimulation by different Hsp70-peptide complexes. The cytotoxicity of such activated PBMCs to different target cells was assayed.

RESULTSThe antigen peptides from different leukaemia cell lines were mixed ones, which could activate PBMC effectively with Hsp70 and stimulate the activated PBMC to proliferate. The proliferative PBMC had specific cytotoxicity to the corresponding leukaemia cells. To Hut-78 cell, Molt-4 cell and Jurkat cell, the cytotoxicity of PBMC activated by either Hut78-peptides or Molt-4-peptides was significantly stronger than that of PBMC activated by HL-60-peptides (P < 0.05). The cytotoxicity to Jurkat cell of PBMC activated by Hut78/Molt-4-peptides was significantly stronger than that of PBMC activated by Hut78-peptides or Molt-4-peptides alone (P < 0.05).

CONCLUSIONAntigen peptide mixture from T lymphocytic leukaemia cells is able to induce specific antitumor immunity. There is a cross-reactivity among antigen peptide mixtures from different T lymphocytic leukaemia cell lines, with the more crossed antigen peptides obtained from the mixtures of different antigen peptides from different T lymphocytic leukaemia cell lines, which suggests that the antigen peptide mixture with broad antigenic spectrum could possibly be prepared by using multiple leukaemia cell lines.

Antigens, Neoplasm ; metabolism ; pharmacology ; Cross Reactions ; HL-60 Cells ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Leukemia, T-Cell ; immunology ; Leukocytes, Mononuclear ; cytology ; drug effects ; Peptides ; pharmacology ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Tumor Cells, Cultured ; chemistry ; immunology

OBJECTIVETo investigate the effects of co-exposure to hyperthermia and lipopolysaccharides (LPS) on tumor necrosis factor-alpha (TNF-alpha) expression in the lungs and small intestines of rats.

METHODSMale pathogen-free Wistar rats were randomly assigned into saline-injected normothermic control (C), saline heat exposure (H), LPS normothermic control (L), and LPS plus heat exposure (HL) groups. The rats in H and HL groups were exposed in a chamber at an ambient dry bulb temperature (Tdb) of 35.0-/+0.5 degrees celsius;, and those in C and L groups to 26-/+0.5 degrees celsius;. In L and HL groups, the rats were given an intravenous injection of LPS 10 mg/kg via the tail vein to induce endotoxemia, and those in C and H group received 10 ml/kg injection. The plasma levels of sTNFrI and sTNFrII were detected at different time points using ELISA. The expression of TNF-alpha in the lungs and small intestines was detected by immunohistochemical SABC method, and the damage of the lungs and small intestines evaluated histologically 120 min after the treatment.

RESULTSCo-exposure to hyperthermia and LPS caused significantly enhanced expressions of TNF-alpha and its receptor sTNFrI and sTNFrII in the plasma and tissues and obvious histopathological damage in the lung and small intestines.

CONCLUSIONCo-stress of hyperthermia and LPS-induced toxicity is associated with the expression of TNF-alpha in the lung and small intestines.

Animals ; Fever ; metabolism ; Hot Temperature ; Immunohistochemistry ; Intestine, Small ; metabolism ; Lipopolysaccharides ; Lung ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Stress, Physiological ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism

Glutamate dehydrogenase (GDH) is a key enzyme in the biosynthesis of glutamate. The GDHs from Corynebacterium glutamicum S9114 the most commonly used strain in glutamate fermentation, were purified and their molecular structures and properties characterized. The coenzymes were also studied in the hope to increase glutamate production. Cells were harvested at mid-exponential phase by centrifugation and washed with Tris-HCl buffer containing DTT and EDTA (pH 7.5). The cells were then disrupted using a French pressure cell press and the supernatant was collected by centrifugation. The extract was concentrated by 70-fold using the AKTA-100 FPLC system employing a DEAE-cellulose ion exchange column, a hydrophobic interaction chromatography (HIC) and Sephadex G-200 gel filtration. The purified extracts contained NADPH-dependent GDH and NADH-dependent GDH. Both of the enzymes were highly specific for the coenzymes. The molecular masses of the NADPH-dependent GDH and its subunit were 188kD and 32kD respectively, suggesting the enzyme is a homo-hexamer. Our data reported for the first time the presence of NADH- dependent GDH in Corynebacterium glutamicum S9114, similar to other microorganisms containing both GDHs. The NADPH-dependent and NADH-dependent GDH in Corynebacterium glutamicum S9114 may participate in the assimilation and dissimilation of ammonia respectively. The absorptions of NADPH-dependent GDH was very weak at 280nm but very high at 215nm, suggesting a low phenylalanine and tyrosine content in the enzyme.
Chromatography, Gel ; Chromatography, Ion Exchange ; Corynebacterium glutamicum ; enzymology ; Glutamate Dehydrogenase ; isolation & purification ; metabolism ; Molecular Weight ; NADP ; metabolism ; Substrate Specificity

The aim of this study was to investigate the expression of integrin linked kinase (ILK) and protein kinase B (PKB/Akt) in acute leukemia (AL), explore the possible effects of ILK/Akt pathway on AL pathogenesis. The expression level of ILK mRNA and Akt mRNA in different types and stages of AL bone marrow mononuclear cells was detected by reverse transcription polymerase chain reaction (RT-PCR). The results showed that the expression of ILK and Akt in do nove AL group was higher than that in AL-CR group and normal control group (P < 0.05), while there was no statistically difference between de nove AL and relapsed AL groups (P > 0.05). The expression of ILK positively correlates with Akt expression in de nove AL group (P < 0.05). It is concluded that the expression of ILK and Akt mRNA is abnormally higher in AL, ILK/Akt pathway may be involved in the pathogenesis and progression of AL and may be a potential therapeutic target for AL.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Female ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics ; Young Adult

Objective To evaluate the characteristics of 99Tcm-ciprofloxacin and explore its feasibility in early detection of secondary infectious foci of severe acute pancreatitis (SAP).Methods Ciprofloxacin was labeled with 99Tcm.The labeling efficiency and radiochemical purity of 99Tcm-ciprofloxacin were calculated and its biodistribution in normal pigs was measured.The recruited baby pigs were divided into three groups:normal control group (6), non-infected group (6) and infected group (16).370-400 MBq of 99Tcm-ciprofloxacin was injected into each pig intravenously.SPECT scanning was performed at 0.5, 1,2, 3, 4 and 6 h after administration.The differences of 99Tcm-ciprofloxacin uptake among groups were calculated and the tracer activity ratio of lesion-to-background was recorded at each time point.The diagnostic value of 99Tcm-ciprofloxacin SPECT imaging for the dectection of secondary infection of SAP was assessed using histopathological results as the gold standard.Variance analysis and least significant difference test were used to analyze the data.Results Both the labehing efficiency and radiochemical purity of 99Tcm-ciprofloxacin were over 90% within 6 h.Organs with rich blood supply, such as kidney, liver and spleen were the target organs for the accumulation of 99Tcm-ciprofloxacin; while no significant uptake was found in gastrointestinal tract or normal pancreas tissue of SAP.Rapid plasma clearance and renal excretion were observed.In the infected group, the lesion was visualized at 1 h after administration.The highest radioactivity ratio of lesion-to-background (3.36 ± 0.33) was at 3 h after administration, which was significantly higher than that of the other time point ( F =99.570, P ＜0.001 ).The sensitivity, specificity, positive and negative predictive values, Youden's index (YI) and Kappa value of 99Tc%ciprofloxacin imaging were 88.2% (15/17), 83.3% (5/6), 93.8% ( 15/16), 71.4% (5/7), 0.715 and 0.667 respectively.Conclusions The biodistribution of99Tcm-ciprofloxacin is suitable for imaging infectious focus of SAP.The optimal imaging time for the detection of secondary infection of SAP is 3 h after administration, with high sensitivity and specificity.

Objective: To investigate the relationship between serum HGF levels and clinical severity of essential hypertension (EH). Methods: The serum HGF concentrations of 44 patients with EH were measur ed by ELISA. Results: The serum HGF levels in patients with EH w ere higher than that in control. Furthermore, the serum HGF levels of EH patient s with coronary atherosclerosis (CAS) were significantly higher than those of EH patients without CAS ［(920.8±250.0） pg/ml vs （747.9±132.1） pg/ml, P <0.01］ or control ［（643.8±98.2） pg/ml, P<0.01)］.The changes of HGF l evel were correlated with the clinical courses (r=0.63, P<0.01) and stag es (r=0.69, P<0.01) of hypertension. Conclusion: HGF may be considered as a new index for the severity of hypertension and an useful bio chemical parameter for estimating the development of atherosclerosis.

OBJECTIVETo investigate the expressions of matrix metalloprotein-2 (MMP-2) and tissue inhibitor of metallopeptidase inhibitor-1 (TIMP-1) in the renal allografts of patients with chronic active antibody-mediated rejection (ABMR), and explore their role in the pathogenesis of ABMR.

METHODSImmunohistochemistry and computer-assisted image analysis were used to detect the expression of MMP-2 and TIMP-1 in the renal allografts of 46 patients with interstitial fibrosis and tubular atrophy (IF/TA), with 15 normal renal tissue specimens as the control. The association of MMP-2 and TIMP-1 with the pathological grade of IF/TA in ABMR was analyzed.

RESULTSThe expressions of MMP-2 and TIMP-1 significantly increased in the renal tissues of the patients as compared with the normal renal tissues (P<0.05). MMP-2 expression tended to decrease, while TIMP-1 and serum creatinine increased with the pathological grades of IF/TA (P<0.05). In IF/TA group, the expression of TIMP-1 was positively correlated to serum creatinine level (r=0.718, P=0.00<0.05).

CONCLUSIONAbnormal expressions of MMP-2 and TIMP-1 can promote the development of renal fibrosis in chronic ABMR.

Adult ; Antibody Formation ; Complement C4b ; metabolism ; Female ; Fibrosis ; etiology ; Graft Rejection ; immunology ; Humans ; Kidney ; metabolism ; Kidney Diseases ; pathology ; Kidney Transplantation ; adverse effects ; immunology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Middle Aged ; Peptide Fragments ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism