AIMTo elucidate the location and effects of transcription factor-nuclear factor-kappaB (NF-kappaB) in lung tissues of rats with chronic obstructive pulmonary disease (COPD).

METHODSFourteen male Wistar rats were randomly divided into COPD model and control groups equally. The COPD model was established by intratracheal instillation of lipopolysaccharide twice and exposure to cigarette smoke daily. We detected the NF-kappaB p65 protein in lung by immunohistochemical method, and the expression of NF-kappaB p65 mRNA in lung by in situ hybridization.

RESULTSImmunohistochemistry, the expression of NF-kappaB p65 protein in alveolar, bronchiolar epithelium and arteriolar endothelium was significantly higher in the COPD group (0.426 +/- 0.007, 0.434 +/- 0.012 and 0.313 +/- 0.007, respectively) than those of the control group (0.115 +/- 0.006, 0.116 +/- 0.005 and 0.095 +/- 0.007, respectively, all P < 0.01). In situ hybridization showed that the expressions of NF-kappaB p65 mRNA in alveolar epithelium (0.203 +/- 0.008), bronchiolar and arteriolar smooth muscle cell (0.208 + 0. 010 and 0.206 + 0.007) of rats in the COPD group were stronger than those in the control group (0.100 +/- 0.006, 0.102 +/- 0.002 and 0.103 +/- 0.003 respectively) by semiquantitative analysis (all P < 0.01).

CONCLUSIONThe expression and nuclear translocation of NF-kappaB may be the basis event of gene expression of many cytokines and inflammatory mediators, which may positively regulate gene expression of many cytokines and inflammatory mediators in various cell lines.

Animals ; Lung ; metabolism ; pathology ; Male ; Pulmonary Disease, Chronic Obstructive ; metabolism ; pathology ; Rats ; Rats, Wistar ; Transcription Factor RelA ; metabolism

Objective To investigate the effect of Geniposide on GLUT 4 expression in insulin -resistance HepG-2 cells.Methods After estab-lish HepG-2 cell model of insulin resistance by high concentrations of insulin-induced in vitro.The cell in model were randomly divided into three groups: control , Geniposide and blank groups.The expression changes of GLUT4 in insulin resistance HepG-2 cells was observe at the time of 4, 12, 24, 36 h after culture by RT -PCR and immunocyto-chemistry.Results GLUT4 expression was first increased and then de-creased with time in HepG -2 cells after treated by Geniposide , and peaked at 8 h.However , compared with the control group , GLUT4 ex-pression was significantly improved in Geniposide group.The difference was statistically significant ( P <0.05 ).Conclusion The improving effect of Geniposide on the improvement of insulin -resistance of HepG-2 cell in vitro is related to the expression of GLUT 4.

Objective To investigate the influence of the physiological conditions and kidney function of the donors on their recipients ’ myco-phenolic acid ( MPA) and mycophenolic acid glucuronide ( MPAG) levels in the early period after renal transplantation.Methods Forty-two pa-tients receiving mycophenolate mofetil ( MMF) and 23 patients receiving enteric-coated mycophenolate sodium ( EC-MPS) , as well as their do-nors were included in this study.For patients who recieving MMF , MPA and MPAG concentrations were detected by HPLC , then AUCMPA and AUCMPAG were calculated by the limited sampling strategy (LSS) model es-tablished by our lab previously.For the patients receiving EC -MPS, AUCMPA and AUCMPAG were calculated by linear trapezoidal rule.Results Spearman test showed significant correlation between recipients ’ AUCMPAG and the age of donors in MMF group , EC-MPS group and the total cohort (P<0.05).Moreover, AUCMPAG in the recipients of elder group (>55 years old) was significantly higher than that in the younger group (≤55 years old) in both MMF group and the total cohort (P<0.05).In addition, AUCMPAG was siginificantly associated with the urea levels of donors before renal transplanta-tion in MMF group and the total cohort ( P<0.05 ).Patients were divided into tacrolimus ( FK506 ) or cyclosporin A ( CsA) group according to their comedication.AUCMPAG was significantly higher in CsA group than that in FK 506 group ( P<0.01 ).AUCMPAG in the recipients of elder group was significantly higher than that in the younger group ( P<0.05), and AUCMPAG was siginificantly associated with the urea levels of donors in FK 506 group (P<0.05).In CsA group, significant correlation between AUC MPAG and the age of donors was showed (P<0.05).Conclusion Our data suggested that the age and the urea level of donors might play roles in the recipients ’ MPAG exposure in the early period after renal transplantation .

OBJECTIVETo explore the association of Chinese medicine constitution susceptibility to diabetic nephropathy (DN) and transforming growth factor (TGF)-β1 (T869C) gene polymorphism.

METHODSTGF-β1 gene polymorphism detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was screened for 180 DN cases and 180 type 2 diabetic mellitus (T2DM) cases without combined DN. Patients with DN were surveyed epidemiologically with constitution in the Chinese medicine questionnaire (CCMQ). Binary logistic regression analysis was utilized to study the correlation between nine types of Chinese medicine constitution and TGF-β1 (T869C) gene polymorphisms.

RESULTSThe DN group has a higher frequency of TGF-β1 (T869C) gene polymorphism than the T2DM group, and CC/CT genotypes than the T2DM group [CC, CT, TT (DN group): 88, 87, 5 (cases) versus (T2DM group) 71, 73, 36 (cases), P<0.05]. The phlegm-dampness constitution, damp-heat constitution, and blood stasis constitution have correlations with TGF-β1 (T869C) gene polymorphism.

CONCLUSIONChinese medicine constitutions were associated with TGF-β1 (T869C) gene polymorphism, a potential predictor of susceptibility to DN in T2DM patients.

Aged ; Body Constitution ; genetics ; Diabetic Nephropathies ; genetics ; Female ; Genetic Predisposition to Disease ; Health Surveys ; Humans ; Logistic Models ; Male ; Medicine, Chinese Traditional ; Polymorphism, Single Nucleotide ; genetics ; Transforming Growth Factor beta1 ; genetics

OBJECTIVESmith-Fineman-Myers syndrome (SFMS) is an X-linked mental retardation syndrome. The authors had ascertained a large Chinese family with SFMS from Shandong and had mapped the disease locus to an interval of 19.8 Mb on Xq25 flanked by markers DXS8064 and DXS8050. Further investigation suggested that SFMS exhibited locus heterogeneity. In this study for facilitating the identification of the gene responsible for SFMS, the additional markers were analyzed to narrow down the candidate region, and four candidate genes (GPC3, MST4,GPCR2 and GLUD2) were chosen and screened for disease-causing mutation.

METHODSPCR and denaturing polyacrylamide gel electrophoresis were used to genotype 13 new polymorphic markers distributed within the candidate region. Mutation detection was accomplished by sequencing the exons and intron-exon junctions of the candidate genes.

RESULTSBy analyzing 13 additional polymorphic markers, SFMS candidate region can be reduced to an interval of 10.18 Mb bounded by XSTR3 and XSTR4, and no disease-causing mutation was identified in the coding regions of four candidate genes.

CONCLUSIONGPCR2 GPC3, MST4 and GLUD2 were excluded as pathogenic genes for SFMS. The refined SFMS locus will assist in the identification and characterization of other candidate genes for SFMS.

Abnormalities, Multiple ; genetics ; Chromosome Mapping ; Chromosomes, Human, X ; Genetic Linkage ; Glutamate Dehydrogenase ; genetics ; Glypicans ; Humans ; Intellectual Disability ; genetics ; Male ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, G-Protein-Coupled ; genetics ; Syndrome

OBJECTIVETo select short tandem repeats(STR) from X chromosome.

METHODSSTR is a universal genetic marker that has changeable polymorphism and stable heredity in human genome. It is a specific DNA segment composed of 2-6 base pairs as its core sequence. It is an ideal DNA marker used in linkage analysis and gene mapping. In this study, 8 short tandem repeats were selected from two genomic clones on X chromosome by using BCM Search Launcher. Primers amplifying the STR loci were designed by using Primer 3.0 according to the unique sequence flanking the STRs. Polymorphisms of the short tandem repeats in Chinese population were evaluated by PCR amplification and PAGE.

RESULTSFive of these STRs were polymorphic. Chi-square test indicated that the distribution of genotypes agreed with Hardy-Weinberg equilibrium (P>0.05).

CONCLUSIONFive polymorphic short tandem repeats have been identified on chromosome X and will be useful for linkage analysis and gene mapping.

Chromosomes, Human, X ; genetics ; Female ; Genotype ; Humans ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

OBJECTIVETo explore the relationship between Chinese medicine (CM) constitutive susceptibility and syndrome diversity in diabetic nephropathy (DN).

METHODSEpidemiologic investigation on constitution adopting the "Constitution in Chinese Medicine Questionnaire" (CCMQ), and survey on syndrome type by CM syndrome scale (preliminary) were carried out in 180 DN patients. Cluster analysis on symptom items was used to determine the syndrome type, and canonical correlation analysis was used to analyze the relationship between patients' constitution and syndrome.

RESULTSBaseline levels in all enrolled patients were not different statistically. Cluster analysis showed 8 syndromes existed in DN patients, namely: I, qi-yin deficiency with qi-stagnancy type; II, yin-yang deficiency with heat-water-blood stasis type; III, qi-yin deficiency with dampness-heat type; IV, yin-yang deficiency with blood-stasis and heat type; V, qi-yin deficiency with stagnant heat type; VI, yin-yang deficiency with inner dampness-heat stagnancy type; VII, yin deficiency with heat stagnancy type; and VIII, Kidney (Shen)-Spleen (Pi) deficiency with stagnant heat type. Correlation analysis on the 8 syndromes and the 9 constitutions showed statistical significant correlations between syndrome III and dampness-heat constitution (P=0.0001); syndrome IV and blood-stasis constitution (P=0.0001); and syndrome VII and yin-deficiency constitution (P=0.0180).

CONCLUSIONCertain relationship revealed between CM constitutions and syndrome types; constitution decides the disease genesis, its syndrome type and prognosis, as well as the change of syndromes.

Aged ; Body Constitution ; Cluster Analysis ; Diabetic Nephropathies ; therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Syndrome

Objective To study the expressed dynamic change and roles of a disintegrin and metalloprotease 9 ( ADAM9) during acute liver injury induced by acetaminophen.Methods Fifty mice were randomly divided into two groups:normal group (n=10) and experiment group (n=40). The mice in experiment group were respectively drawn blood by removing the eyeballs and serum was separated to detect the activity of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) at 6, 24, 42 and 72 h after intraperitoneal injection of acetaminophen 550 mg? kg-1 .The activity of serum AST and ALT in the mice of normal group were also detected.The expression of hepatic ADAM9 at protein and mRNA levels were detected by western blot and RT -PCR method in the mice of normal group and experiment group at different time points after acetaminophen injection.Results The expression of ADAM9 protein and mRNA were significantly down -regulated at 6 h ( P<0.05) , and the expression of ADAM9 protein and mRNA reached the lowest level at 24 h ( P <0.05 ) . After that, the expression of ADAM9 protein and mRNA were gradually increased.The expression of ADAM9 was significantly increased at 42 h (P<0.05),and then recovered closed to the normal level at 72 h after acetaminophen injection.Conclusion ADAM9 is remarkably differently ex-pressed during acute liver injury induced by acetaminophen, which indicates that ADAM9 may play an important pro-tective role during acute liver injury induce by acetaminophen in mice.

Objective To investigate effect of snail polypeptide on skin healing and expression of basic fibroblast growth factor ( bFGF) mRNA in rats with scalded wound.Methods The deepⅡscald model was estab-lished in 60 rats by using 90 ℃ hot water.And the rats were randomly divided into model group and the test group.Rat burn sites in test group were smeared with SP solution(10 mg? mL-1 ).Skin healing rate,skin healing time and the scab off time were observed in each group of rats. The mRNA expression of skin tissue bFGF was detected by RT-PCR. Results Healing rate of test group at different times after treatment were significantly higher than the model group, with statistically significant difference ( P <0.05 or P <0.01 ) .Compared with the model group, burn the skin healing time scab off times are significantly shorter in test group, the difference was statistically significant ( P <0.05 ) .Mean-while, SP increased the expression of bFGF mRNA at different times. Conclusion SP can promote wound healing in rat skin burns, and its mechanism may be related to regulating bFGF mRNA expression.

Objective To study the effects of melatonin ( MT) on insulin signaling transduction defect induced by linoleic acid ( LA) and the un-derlying mechanism.Methods HepG2 cells were divided into BSA group, LA group, LA+MT group and LA+VitC group cultured with va-rious agent concentrations of 10% BSA, or 0.5 mmol? L-1 LA, or 10μmol? L-1 MT plus 0.5 mmol? L-1 LA, and 1 mmol? L-1 Vitamin C plus 0.5 mmol? L-1 LA, respectively for 24h.The level of intracellular ROS was measured by fluorometry.Phosphorylation levels of protein ki-nase B ( PKB) , Forkhead box O1 ( FoxO1 ) , and c-Jun N-terminal kinase ( JNK) were determined in total cell lysates by Western-blotting with insulin stimulation.Results ROS level increased significantly in LA group compared with that in BSA group, and was significantly higher compared with that in LA +MT group ( P <0.05 ) . There was no significant difference in ROS level between LA+MT group and BSA group.Compared with LA group, the production of ROS in LA+VitC group decreased, but significantly higher than that in LA+MT group ( P<0.05 ) .Compared with BSA group, the phosphorylation levels of both PKB and FoxO1 in LA group significantly decreased, whereas, phospho-rylation level of JNK increased significantly ( P<0.05 ) .Copared with LA group, phosphorylation levels of PKB and FoxO1 in LA +MT group increased significantly, whereas, phosphorylation level of JNK significantly decreased (P<0.05).Conclusion LA causes the increased production of ROS in HepG2 cells.MT treatment can clear the excessive LA-induced ROS, reduce the activation of c-Jun amino terminal kinase and repair the impaired insulin downstream signal.