Mucin as an important member in gastrointestinal mucus layer has become the focus of research. Many studies suggested that MUC1 mucin is involved in the mechanism of Helicobacter pylori( Hp)infection. It is important to understand the relationship between MUC1 mucin and Hp for preventing Hp infection and gastric cancer. This article reviewed the relationship between MUC1 mucin and Hp.

To study the effect of genistein on the proliferation of cardiac fibroblasts(CF), CFs were cultured from neonatal rat hearts, DNA synthesis of the cells was determined by incorporation of ［3H］TdR into DNA, the cell cycle was measured by flow cytometric analysis. Genistein(0.5－50 μmol*L-1) attenuated 2.5% fetal calf serum-induced proliferation of CF in concentration-dependent manner. Genistein(50 μmol*L-1) arrested CF cell progression at G2/M phase. The results suggest that genistein be a potential substance for treatment of cardiac fibrosis.

Objective To explore the expressions and significance of miRNA - 101,enhancer of ZESTE homo-log 2(EZH2)and transforming growth factor(TGF)- β1 in the kidneys with complete uniateral ureteral obstruction (CUUO). Methods Thirty male SD rats,(6 ± 1)weeks old,weighted(150 ± 10)g,were divided into sham group, 7 - day group with CUUO and 14 - day group with CUUO by using random number table method,10 rats in each group. The obstructed kidney samples were collected in 7 and 14 days,respectively,for detecting the expression of miRNA -101 by real time - polymerase chain reaction(RT - PCR)and TGF - β1 and EZH2 protein by Western blot,immuno-histochemistry and hematoxylin - eosin staining. Their correlated expressions were analyzed. Results RT - PCR results showed that the expressions of miRNA - 101 in sham group and 7 - day group with CUUO were(12. 69 ± 1. 60)times and(3. 74 ± 1. 24)times which were higher than those of 14 - day group with CUUO,respectively,there was a signifi-cant difference among these 3 groups(P ﹤ 0. 05). The expressions of TGF - β1 and EZH2 proteins were 1. 14 ± 0. 12, 1. 04 ± 0. 14,0. 76 ± 0. 18 and 1. 04 ± 0. 04,0. 89 ± 0. 03,0. 73 ± 0. 02 in 14 - day group with CUUO,7 - day group with CUUO and sham group,respectively. There was a negative correlation between miRNA - 101 with EZH2( r =- 0. 92,P ﹤ 0. 05),and negative correlation with TGF - β1(r = - 0. 63,P ﹤ 0. 05),and positive correlation between EZH2 and TGF - β1(r = 0. 67,P ﹤ 0. 05);the expressions of miRNA - 101,EZH2 and TGF - β1 were associated with each other in obstruction renal in different time periods. Conclusions With the extension of obstruction time, miRNA - 101 expression decreased,EZH2 and TGF - β1 expression increased,evidently,which indicates that the de-velopment of renal interstitial fibrosis may be affected through regulating EZH2 expression of renal obstruction through miRNA - 101 in the young rats.

Objective: To investigate the effects of tumor necrosis factor-α (TNF-α) monoclonal anti-body on nuclear factor-κB (NF-κB) activation and inducible nitric oxide synthase (iNOS) expression in rats with pulmonary fibrosis induced by silica dust. Methods: A total of 48 male Wistar rats were randomly divided into intervention group, silica dust exposure group, and control group, with 16 rats in each group. The rats in the intervention group were given intratracheal injection of 50 mg silicon dioxide dust once to establish a rat model and then treated with subcutaneously injected TNF-α monoclonal antibody 15 mg/kg for 5 consecutive days at 2-6 days after the establishment of the model. The rats in the silica dust exposure group were treated with the same method to establish the model and then given subcutaneous injection of the same volume of normal saline. The rats in the control group were given intratracheal and subcutaneous injection of normal saline. In both groups, 8 rats each were sacrificed at 7 and 14 days after the establishment of the model. Hematoxylin-eosin staining or Masson staining was used to observe morphological changes in lung tissue, ELISA was used to measure the serum level of TNF-α, IHC was used to measure the expression of NF-κBp65 in lung tissue, Western blot was used to measure the protein expression of I-κB in lung tissue, and RT-qPCR was used to measure the transcriptional level of iNOS mRNA in lung tissue. Results: Compared with the control group, the silica dust exposure group had significant increases in the lung inflammation score (3.375±1.061 and 2.500±0.535) , serum TNF-α level (86.405±20.494 and 77.064±11.829) , absorbance of cells with positive NF-κBp65 in lung tissue (0.297±0.05 and 0.287±0.039) , and mRNA expression of iNOS (12.906±0.590 and 12.600±0.517) at 7 and 14 days after dust exposure, a significant increase in pulmonary fibrosis score at 14 days (3.250±0.707) , and significant reductions in the protein expression of I-κB at 7 and 14 days (0.579±0.141 and 0.748±0.081) (P<0.05) . Compared with the silica dust exposure group, the intervention group had significant reductions in the lung inflammation score at 7 days (2.375±1.061) , pulmonary fibrosis score at 14 days (2.375±1.061) , serum level of TNF-α at 7 and 14 days (66.565±19.850 and 58.734±16.335) , absorbance of cells with positive NF-κBp65 in lung tissue at 7 and 14 days (0.248±0.028 and 0.238±0.027) , and mRNA expression of iNOS at 7 and 14 days (11.656±0.405 and 12.025±0.618) , as well as significant increases in the protein expression of I-κB at 7 and 14 days (0.802±0.165 and 0.888±0.144) (P<0.05) . Conclusion TNF-α monoclonal antibody can inhibit the activation of the NF-κB signaling pathway and down-regulate the expression of iNOS, and thus exerts a certain protective effect on lung tissue in rats with pulmonary fibrosis induced by silica dust.

BACKGROUNDNF-kappaB p65 was shown to inhibit transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme in gluconeogenesis in the liver. To understand the mechanism of action of NF-kappaB p65, we investigated the nuclear receptor corepressor in the regulation of PEPCK transcription.

METHODSRat H4IIE cells, human hepatoma HepG2 cells and human embryo kidney (HEK) 293 cells were used in this study. The transcriptional activity of a rat PEPCK gene promoter (-490/+100) was analyzed in HepG2 cells, a HepG2 super suppressor IkBalpha (ssIkBalpha) stable cell line, and HEK 293 cells. The effects of p65 and ssIkBalpha on a rat PEPCK gene promoter were observed using the PEPCK luciferase reporter system. The interaction of the cAMP-response- element-binding (CREB) protein, histone deacetylase 3 (HDAC3) and silencing mediator for retinoic and thyroid hormone receptors (SMRT) with the PEPCK gene promoter were investigated using the chromatin immunoprecipitation (ChIP) assay. p65 cotransfection and RNAi-mediated gene knockdown were used to determine the corepressor involved in the inhibition of PEPCK by NF-kappaB p65 and the transcriptional regulation of CREB by NF-kappaB p65.

RESULTSNF-kappaB p65 inhibited PEPCK expression and the inhibition was blocked by ssIkBalpha. The inhibitory effect of p65 was completely blocked in a HepG2 stable cell line in which ssIkBalpha was expressed. HDAC3 or SMRT knockdown led to a significant up-regulation of PEPCK reporter activity in the presence of p65 cotransfection. In the ChIP assay the interaction of HDAC3 and SMRT with the PEPCK gene promoter was induced by p65 activation, but the CREB signal was reduced. Transcriptional activity of CREB was inhibited by NF-kappaB p65 cotransfection. The inhibitory effect of NF-kappaB p65 was blocked by HDAC3 RNAi or SMRT RNAi.

CONCLUSIONSThe study showed that the inhibition of PEPCK by NF-kappaB p65 was dependent on HDAC3 and SMRT, which form a nuclear corepressor complex for transcriptional inhibition. The transcription factors NF-kappaB p65 and CREB share the same corepressor HDAC3-SMRT, and the corepressor exchange leads to inhibition of PEPCK gene transcription by NF-kappaB p65.

Animals ; Blotting, Western ; Cell Line ; Chromatin Immunoprecipitation ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Hep G2 Cells ; Histone Deacetylases ; genetics ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Nuclear Receptor Co-Repressor 2 ; genetics ; metabolism ; Phosphoenolpyruvate Carboxykinase (ATP) ; genetics ; Promoter Regions, Genetic ; genetics ; Protein Binding ; genetics ; physiology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor RelA ; genetics ; metabolism

OBJECTIVETo explore the regulatory effect of arginine on the secretion of hepatic insulin-like growth factor-1 (IGF-I), and the mechanism of enhancing the immune function by arginine.

METHODSWistar rats were randomly divided into normal control (NC), wound control (WC), and wound with arginine (Arg) groups, with 8 rats in each group. The rats in WC and Arg groups were inflicted with soft tissue trauma on the back. The rats in Arg group were fed a diet supplemented with 5% arginine for one week, while those in NC and WC groups were fed with glycine. The serum contents of arginine, ornithine, growth factor (GH), NO and IGF-I were determined 7 days after feeding. T cell proliferation and IGF-I mRNA expression in hepatic tissue were also measured. Meanwhile, the rat hepatocytes were cultured in serum-free medium containing different concentrations of arginine. The supernatant was collected for the determination of IGF-I level.

RESULTS1). There was no obvious difference of the serum level of arginine and ornithine between NC and WC groups (P > 0.05), but the contents of them were obviously higher in the Arg group compared with other two groups (P < 0.01). 2). No difference in the serum GH level was found among all the groups (P > 0.05), but the serum NO content in WC and Arg groups was significantly lower than that in NC group (P < 0.01), and the serum IGF-I content in WC group decreased obviously compared with that in NC group (P < 0.01). 3). The thymocyte proliferation rate in WC group was also markedly lower than that in NC group (P < 0.01), but that in Arg group was improved compared with WC group (P < 0.01). 4). The expression of hepatic IGF-I mRNA: The relative value of IGF-I mRNA was 1.19 +/- 0.06, 1.08 +/- 0.06 and 1.29 +/- 0.06 in NC, WC and Arg, respectively, while the value in WC was lower than that in NC (P < 0.05) group, and that in Arg group was much higher than that in WC group (P < 0.01). 5). The IGF-I level in the supernatant of cultured hepatocytes: When Arg concentration was 0.0750, 0.7500, 7.5000 mmol/L in the culture medium, the IGF-I level in the supernatant of hepatic cell medi-um was obviously higher than that in the medium without arginine (P < 0.01). Although IGF-I level decreased in the culture medium with arginine in the dose of 37.5000 mmol/L, it was still obviously higher than that in the medium without arginine (P < 0.01).

CONCLUSIONArginine could also produce the immune enhancing effect by stimulating hepatic IGF-I secretion.

Animals ; Arginine ; pharmacology ; Enteral Nutrition ; Insulin-Like Growth Factor I ; metabolism ; Liver ; drug effects ; secretion ; Male ; Rats ; Rats, Wistar ; Soft Tissue Injuries ; metabolism ; therapy

Objective To explore the significance of urine deoxypyridinoline to diagnosis on osseous metastasis of lung neoplasms.Methods.182 cases with lung carcinoma was divided into two groups.One group was case with osseous metastasis,the other group was case without osseous metastasis,uDPD/uCr, uCa/Cr,sCa and sAKP in two groups were respectively compared.Sensitivity and specificity of these indexes to diagnosis on osseous metastasis of lung cancer were also acalculated and compared.80 cases without osseous metastasis were follow-up for 6 months.Results The ratio of uDPD/uCr with osseous metastasis group[(12.35?2.65)nmol/mmol]was significantly higher than that of without osseous metastasis group [(7.76?2.11)nmol/mmol](t=2.46,P

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

OBJECTIVE: To introduce the design of customized pelvic prosthesis, to evaluate the biomechanical property under three load conditions of customized pelvic prosthesis under three load cinditions. METHODS: A titanium alloy prosthesis for reconstruction of pelvic tumors was designed by CAD software. The strength and stiffness of the custom prosthesis under static and slow gait conditions were analyzed and evaluated by finite element method. RESULTS: The results of the finite element analysis suggested that the maximum von Mises stress in the pelvic under three load conditions were 39.0, 202.8 and 42.4 MPa; the maximum displacement were 0.199, 0.766 and 0.847 mm. The maximum von Mises stress in the prosthesis under three load conditions were 62.3, 318 and 468 MPa. The maximum Von Mises stress in the Ti-alloy prosthesis and pelvic was far smaller than the yield strength of Ti-alloy. CONCLUSIONS The study can design the size and shape of prosthesis accurately according to patient's condition. The finite element method can reduce the bone stress level and fracture risk, prolong the service life of prostheses, and ensure the safety and stability of the postoperative patients under normal gait.
Biomechanical Phenomena ; Finite Element Analysis ; Fractures, Bone ; Gait ; Humans ; Prostheses and Implants ; Prosthesis Design ; Stress, Mechanical

Objective To explore the effect of chlorpromazine on neocortical high-voltage spindles(HVSs)and facial twitching in freely moving normal adult rats.Methods Continuous video,neocortical electroencephalogram(EEG)and facial electromyography(EMG)were recorded simultaneously for at least 6 h in freely moving rats.The number and mean duration of HVSs,and the facial muscular activity were analyzed after administration of chlorpromazine.Results HVSs spontaneously appeared in behavioral waking-immobility rats with accidental hyperspasmia.After rats being administrated with chlorpromazine at a dose of 1.5(mg/kg)and 3.0(mg/kg),the facial rhythmic muscular activity increased,the duration of rhythmic muscular activity prolonged,the number of HVSs increased and the mean duration of HVSs prolonged.Conclusion Chlorpromazine could enhance the physiological HVSs and facial twitching,indicating that the side effect at the extrapyramidal system of antipsychotic maybe relate to HVSs and dopamine receptor may contribute to the regulation of HVSs.