plasmid can be used to study the pathogenic mechanism of target gene products of L.interrogans.

Objective To evaluate the effects of sevoflurane postconditioning on the oncosis and apoptosis in cardiomyocytes during ischemia-reperfusion (I/R) in isolated rat hearts and the role of extracellular signalregulated protein kinase 1/2 (ERK1/2) signal transduction pathway in it.Methods Seventy-two isolated rat hearts perfused in a Langendorff apparatus were randomly divided into 6 groups (n =12 each) using a random number table:sham operation group (group S),myocardial I/R group (group I/R),sevoflurane postconditioning group (group SP),PD98059 vehicle dimethyl sulfoxide (DMSO) group (group DMSO),selective ERK1/2 inhibitor PD98059 group (group PD),and sevoflurane postconditioning + PD98059 group (group SP + PD).The hearts were subjected to ischemia for 30 min followed by 2 h reperfusion in the other groups except group S.In SP,DMSO and PD groups,the hearts were perfused with K-H solution saturated with 3.0％ sevoflurane,DMSO (＜0.2％) and PD98059 (20 μmol/L),respectively,for 15 min starting from the end of ischemia until 15 min of reperfusion,and then with plain K-H solution for 105 min.In group SP+ PD,the hearts were perfused with K-H solution saturated with 3.0％ sevoflurane and PD98059 for 15 min starting from the end of ischemia until 15 min of reperfusion.Myocardial infarct size and expression of porimin and caspase-8 proteins (by Western blot) were measured at the end of reperfusion.Results Compared with S group,the myocardial infarct size was significantly increased,and the expression of porimin and caspase-8 proteins was up-regulated in the other groups (P ＜ 0.05).Compared with I/R group,the myocardial infarct size was significantly decreased,and the expression of porimin and caspase-8 proteins was down-regulated in group SP (P ＜ 0.05),and no significant changes were found in the other groups (P ＞ 0.05).Conclusion Sevoflurane postconditioning can activate ERK1/2 signal transduction pathway and inhibit the oncosis and apoptosis in cardiomyocytes,thus attenuating I/R injury in isolated rat hearts.

Objective To evaluate the effects of sevoflurane postconditioning on the expression of 70 kD aribosomalprotein S6 kinase (p70S6K) during ischemia-reperfusion (I/R) in isolated rat hearts.Methods Pathogen-free male Sprague-Dawley rats,aged 3 months,weighing 270-350 g,were used in the study.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95 ％ O2-5 ％ CO2.Ninety isolated rat hearts with I/R injury were randomly divided into 3 groups (n =30 each):sham operation group (group S),I/R group (group I/R),and sevoflurane postconditioning group (group SP).The hearts were subjected to ischemia for 30 min followed by 2 h reperfusion.In group SP,the hearts were perfused with K-H solution saturated with 3.0％ sevoflurane for 15 min starting from the end of ischemia until 15 min of reperfusion,and then with plain K-H solution for 105 min.At 2 h of reperfusion,myocardial infarct size was measured,the percentage of myocardial infarct size was calculated,and the phosphorylated p70S6K (p-p70S6K)/total p70S6K (tp70S6K) ratio,and cytoplasm,cytochrome C,and caspase-8 expression was measured.Results Compared with group S,the percentage of myocardial infarct size and p-p70S6K/t-p70S6K ratio were significantly increased,the expression of cytochrome C,and caspase-8 was up-regulated,and the expression of cytochrome C was downregulated in I/R group.Compared with I/R group,the percentage of myocardial infarct size was significantly decreased,the ratio of p-p70S6K/t-p70S6K was increased and cytochrome C expression was up-regulated,and the expression of cytochrome C and caspase-8 was down-regulated in SP group.Conclusion Sevoflurane postconditioning can mitigate I/R injury to isolated rat hearts,and up-regulation of p-p70S6K expression,inhibition of transfer of cytochrome C from mitochondria to cytoplasm,and reduced cell apoptosis are involved in the mechanism.

Objective To evaluate the role of the mitochondrial ATP-sensitive potassium (mito-KATP)channel in sevoflurane preconditioning-induced delayed cardioprotection against ischemia-reperfusion (I/R) injury in isolated rat hearts.Methods Seventy-two adult male Sprague-Dawley rats were randomly divided into 6 groups (n =12 each):control group (group CON),I/R group,sevoflurane control group (group SEVO),sevoflurane preconditioning group (group SWO P),5-hydroxydeconoate (5-HD) + sevoflurane preconditioning group (group 5-HD+ SWOP) and 5-HD control group (group 5-HD).The rats were exposed to 33％ pure oxygen for 1 h in groups CON and I/R.The rats were exposed to 2.5％ sevoflurane for 1 h in groups SEVO and SWOP.5-HD (a mito-KATP channel inhibitor) 10 mg/kg was injected intraperitoneally 30 min before sevoflurane preconditioning in group 5-HD + SWOP.5-HD 10 mg/kg was injected intraperitoneally in group 5-HD.The hearts were immediately removed and perfused in a Langendorff apparatus.The hearts were made globally ischemic for 30 min followed by 120 min reperfusion in groups I/R,SWOP,5-HD + SWOP and 5-HD.The expression of phosphorylated protein kinase C-epsilon (p-PKC-ε) and caspase-8 was measured by Western blot immediately before ischemia (T0) and at 120 min of reperfusion (T1).The myocardial infarct volume was measured by TTC staining.Results Compared with group CON,the myocardial infarct volume was significantly increased at T1 in groups I/R,SWOP,5-HD +SWOP and 5-HD,p-PKC-ε expression was up-regulated at T0 in groups SEVO and SWOP and at T1 in groups I/R,SWOP,5-HD + SWOP and 5-HD,and caspase-8 expression was down-regulated at T1 in group SEVO (P ＜0.05).Compared with group I/R,the myocardial infarct volume was significantly decreased at T1 in groups SWOP and 5-HD + SWOP,p-PKC-ε expression was up-regulated at T0 in groups SEVO and SWOP,and caspase-8 expression was down-regulated at T1 in group SWOP (P ＜ 0.05).Compared with group SWOP,the myocardial infarct volume was significantly increased,p-PKC-ε expression was down-regulated at T0,and caspase-8 expression was up-regulated at T1 in group 5-HD + SWOP (P ＜ 0.05).Conclusion The mito-KATP channel is involved in sevoflurane preconditioning-induced delayed cardioprotection against I/R injury in isolated rat hearts through upregulation of p-PKC-ε expression before ischemia and inhibition of cell apoptosis during reperfusion.

Objective To investigate the immune response of the fusions of ovarian carcinoma cells to dendritic cell(DC) transfected with interleukin(IL)-12 gene in vitro. Methods Recombinant human granulocyte macrophage colony stimulating factor and recombinant human tumor necrosis factor ? were used to generate DC from cord blood in vitro. IL-12 fusion gene was cloned into a eukaryotic vector-pcDNA3.1(+). DC were transfected with IL-12-pcDNA3.1(+) using lipofectamine transfection method and electric transfection method respectively. Then polyethylene glycol mediated the fusion of transfected DC and ovarian carcinoma cells, and 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test was carried out to observe the immune response. Results DC were generated in vitro from cord blood and expressed high levels of costimulatory(50%) and MHC Ⅱ molecules(99%). RT-PCR showed that IL-12-pcDNA3.1(+) had been successfully transfected into DC. RT-PCR and immunofluorescence analysis showed that DC were fused with ovarian carcinoma cells successfully. Then the fusion cells of ovarian carcinoma cells to transfected DC, and the fusions of ovarian carcinoma cells to DC were cocultured with peripheral blood mononuclear cell respectively. MTT test showed that both fusions could induce cytolytic T cell activity and lysis of ovarian carcinoma cells,and the former effect was stronger. Conclusion The cytolytic T cell activity induced by the fusions of ovarian carcinoma cells to transfected DC is enhanced.

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.

Marine Actinobacteria are emerging as new resources for bioactive natural products with promise in novel drug discovery. In recent years, the richness and diversity of marine Actinobacteria from the South China Sea and their ability in producing bioactive products have been investigated. The objective of this work is to isolate and identify bioactive secondary metabolites from a marine actinobacterium SCSIO 1934 derived from sediments of South China Sea. The strain was identified as a Streptomyces spieces by analyzing its 16S rDNA sequence. Streptomyces sp. SCSIO 1934 was fermented under optimized conditions and seven bioactive secondary metabolites were isolated and purified by chromatographic methods including colum chromatography over silica gel and Sephadex LH-20. Their structures were elucidated as 17-O-demethylgeldanamycin (1), lebstatin (2), 17-O-demethyllebstatin (3), nigericin (4), nigericin sodium salt (5), abierixin (6), respectively, by detailed NMR spectroscopic data (1H, 13C, COSY, HSQC and HMBC). This work provided a new marine actinobacterium Streptomyces sp. SCSIO 1934, capable of producing diverse bioactive natural products.
Anti-Bacterial Agents ; chemistry ; China ; DNA, Ribosomal ; chemistry ; genetics ; Geologic Sediments ; microbiology ; Oceans and Seas ; RNA, Ribosomal, 16S ; genetics ; Streptomyces ; chemistry ; classification ; genetics ; isolation & purification

Objective To evaluate the effectiveness of a rat model for comorbidity of Tourette syndrome and anxiety with empty water bottle stimulation plus iminoodipropionitrile(IDPN) injection.Methods The 48 male SD rats were randomly divided into four groups:the blank control group,the TS group,the anxiety group and the comorbidity group.The blank control group was injected with saline for 7 days.The TS groop was injected with 3,3-iminodipropionitrile (IDPN) with 250 mg/kg once a day for 7 consecutive days.The anxiety group was given empty water bottle stimulation for 21 consecutive days.The comorbidity group was given empty water bottle stimulation plus IDPN injection.At the end of the 3rd week,the behavioral changes of the stereotyped movement,elevated plus-maze and open field of the rats in each group were measured,and the contents of monoamine neurotransmitters in striatum and hippocampus were determined by HPLC.Results The results of stereotyped movement showed that there was no significant difference between the groups except for the blank control group.The elevated plus-maze test showed that the 0E/TE values of the comorbidity group (21.33±11.35) ％ and the anxiety group (17.68±16.89) ％ were significantly decreased,lower than that of the blank control group (73.24± 19.33) ％ and TS group(61.43±21.84) ％.The results of open field test showed that the total scores of open field in the comorbidity group(15.22±9.87)and anxiety group (11.17±10.76) were lower than that of the blank control group (41.86±33.30) and TS group(48.83± 17.65) (P＜0.01).However,there was no significant difference between the comorbidity group and the anxiety group.The test of monoamine neurotransmitters in striatum showed that the content of HIAA in the comorbidity group(0.03±0.00) ng/mg was the highest,and that of the TS group and anxiety group (0.02±0.00) ng/mg was higher than that of the blank control group (0.01±0.00) ng/mg (P＜0.01).The DA test showed that the content of DA in the comorbidity group (0.03±0.00) ng/mg was the highest,and that of the comorbidity group,TS group(0.02±0.00) ng/mg and anxiety group was higher than that of the blank control group(0.01±0.00) ng/mg (P＜0.01).The expression of 5-HT was most significant among the groups (P＜0.01),and there was significant difference between the anxiety group ((0.011 ± 0.001) ng/mg)and the comorbidity group ((0.014±0.002) ng/mg) (P＜0.01).The expression of HVA in the three model groups ((0.05±0.00) ng/mg) was higher than that in the blank group ((0.02±0.00) ng/mg) (P＜ 0.01).The expression of DOPAC in the TS group ((0.23±0.02) ng/mg) was higher than that in the blank control group((0.16±0.01) ng/mg) and comorbidity group ((0.16±0.02) ng/mg) (P＜0.01).The test of monoamine neurotransmitters in hippocampus showed that the content of 5-HT in the comorbidity group ((0.14±0.02) ng/mg) was the highest,followed by the anxiety group ((0.1 ± 0.03) ng/mg) and the TS group ((0.07±0.04) ng/mg),which were all higher than the blank control group((0.04±0.03) ng/mg) (P＜0.05,P＜0.01),and there were significant differences between the comorbidity group and the TS group or anxiety group (P＜0.01).The expressions of HIAA and HVA were higher in the comorbidity group((0.44±0.04)ng/mg,(0.01±0.00) ng/mg),TS group ((0.46±0.15) ng/mg,(0.01 ±0.01) ng/mg) and anxiety group ((0.46±0.08)ng/mg,(0.01±0.00) ng/mg) than that in the blank control group((0.21±0.10)ng/mg,(0±0) ng/mg) (P＜0.05,P＜0.01).Conclusion This study confirms the reliability of the model and it is an ideal animal model for the study of TS with comorbidity of anxiety,which can be used for follow-up research.

Objective With multi?center investigation, to assess the life quality of patients with maintained hemodialysis (MHD) in Liaoning Province and to explore the relationship among the mineral metabolism, the life quality of the patients with MHD, and the repeated hospitalization within the latest three years. Methods 1192 patients with hemodialysis (at least 3 months) from January to March in 2015 at ten blood purification centers in Liaoning Province were selected for the cross?————————sectional survey. The Kidney Health?related Quality of Life (HRQOL) version 1.3 was used to evaluate the MHD patients' life quality. The total length of hospitalization was divided into four groups: 0 days, 3 to 15 days, 16 to 30 days and above 30 days. Results When serum calcium value ranged from 2.1 to 2.5 mmol/L, kidney?disease component summary (KDCS), mental component summary (MCS), physical component summary (PCS) and SF?36+KDCS corresponded to a higher value (P<0.05). When serum phosphorus value ranged from 1.13 to 1.78 mmol/L, KDCS and SF?36+KDCS corresponded to a higher value (P<0.05). When the calcium phosphorus product value ranged from 40.68 to 49.94, MCS corresponded to a higher value (P<0.05). KDCS showed a linear correlation with age (P<0.001), dialysis age, serum calcium (less than or equal to 2.5 mmol/L) (P<0.05); PCS showed a linear correlation with age (P<0.001) and dialysis age (P<0.05); SF?36+KDCS showed a linear correlation with age (P<0.001), and serum calcium (less than or equal to 2.5 mmol/L) (P<0.05), while age and dialysis age were negatively correlated. The hospitalization days showed a linear correlation with age, dialysis age (P<0.001) and serum phosphorus, calcium phosphorus product value (P<0.05), while dialysis age and calcium phosphorus product value were negatively correlated. Among different groups of total hospitalization days in three years, age, hemodialysis age, serum calcium, serum phosphorus, calcium?phosphorus product value and quality of life values were all statistically significant (P<0.05). Conclusions The life quality of patients with MHD were correlated with serum calcium, phosphorus, calcium and phosphorus product value, iPTH, dialysis age and age, while age and dialysis age were of negative correlation. The total number of hospitalization days in 3 years was closely linearly correlated with age and dialysis age, significantly correlated with serum phosphorus, calcium and phosphorus product value, while dialysis age, calcium and phosphorus product value were in a negative correlation. The total number of hospitalization in 3 years was correlated with the patients' age, dialysis age, serum calcium, serum phosphorus, calcium and phosphorus product value and quality of life.

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