Objective To investigate the influence of plasmid spread and ampD mutation to Enterobacter cloacae that leads to the AmpC β-lactamase change from inducible type to constitutive type. Methods The Enterobacter cloacae were isolated from the patients with nosocomial infection. The inducible type isolations and their constitutive type changers were put into the same group. The plasmid ampC gene and chromatin ampD gene in pairs in each group were amplified, sequenced and compared. Results Of 195 patients infected by Enterobacter cloacae of inducible type, 25 (12.82%) were changed to the ones of constitutive high type. In these 25 changed groups, 10 were caused by plasmid spread, 10 by ampD mutation, 1 by both, and 4 by neither. Twelve changed constitutive type strains had ampD significant mutations, in which 7 were frame-shift mutations and 5 were spot mutations. Conclusions The change ratio of Enterobacter cloacae from inducible type to constitutive type is rather high. Both plasmid spread and ampD mutation are possibly the mechanism of such change. Plasmid mediated AmpC β-lactamase spreads among different species and interregionally. The mutation rate of chromatin ampD gene is also higher than the natural mutation rate. These two mechanisms should be considered in clinical treatment.

Some intracellular enzyme-activities of BCG-activated alveolar macrophages (AM) and peritoneal macrophages (PM) were studied. It was found that (in normal) the activities of acid phosphatas (ACP), succinate dehydrogenase (SDH), non-specific esterase (NSE) in AM were higher than those in PM, but that of lactate dehydrogenase (LDH) was lower. Both in BCG activated AM and PM, the activities of ACP, SDH, NSE increased significantly, and that of LDH also enhanced in activated AM but not in PM.

Objective To reveal the influence factor of ampD specific mutation on the transformation of AmpC β-lactamase from non-high-productive type to derepressed the high-productive type in Enterobacter cloacae. Methods The selected cbromatin-mediated derepressed high-productive strains of Enterobacter cloacae and genomic DNA were extracted, and genomic ampD was amplified by using PCR, and then sequenced and confirmed by specific mutation. Use site-directed mutagenesis to mutate the wild-type Enterobacter cloacae for the above site. The changes of AmpCβ-lactamase types were detected by the improved Cefotaxime three-dimensional test. Results Of the 121 strains of Enterobacter cloacae,15 strains were ehromatin-mediated derepressed high-productive type,8 strains had significant mutations and 7 strains of site-directed mutagenesis were established. Ecl MA(274 inser-ting A)、Ecl MC(327 missing C)and Ecl MF(27 inserting G)were changed from non-high-productive type to derepressed-high-pro-ductive type,but Ecl MB(371 inserting T), Ecl MD(515 missing C), Ecl ME(324 C→A), Ecl MG(238C→A 302T→A)did not change in AmpC β-lactamase types. Conclusion The significant mutations that could lead to the transformation of AmpC β-lacta-mase were the Frame-shift mutations that could create the change of large region of amino acid. However, other mechanisms for the derepressed high-productive AmpC β-lactamase were not excluded in the study.

Objective To denoise digital radiographic images well.Methods A technique was presented that used the Anscombe's transformation to adjust the original image to a Gaussian noise model based upon the wavelet denoising method and the wavelet-domain Hidden Markov Tree(HMT) model.Wavelet domain HMT models were used to determine the dependencies of multiscale wavelet coefficients through the state probabilities of the wavelet coefficients,whose sedistribution densities could be approximated by Gaussian mixture model.Results The proposed method could keep natural images edges from damaging and increase PSNR.Conclusion Quantitative and qualitative DR images assessment shows that the proposed algorithm outperforms the traditional Gaussian filter in terms of noise reduction,quality of details and bone sharpness.

Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Calbindin 2 ; Cell Adhesion Molecules ; metabolism ; Claudins ; metabolism ; Diagnosis, Differential ; Epithelial Cell Adhesion Molecule ; Humans ; Matrix Metalloproteinases ; metabolism ; Mucin-1 ; metabolism ; Pleural Effusion, Malignant ; diagnosis ; metabolism ; Receptor, trkA ; metabolism ; S100 Calcium Binding Protein G ; metabolism

The producing condition and properties of chitinase secreted by Nomuraea rileyi strain CQ031021 were studied.The optimal conditions for the strain to produce chitinase are 6 days of 28℃ with the initial pH 6.0,and the liquid medium containing 2.0% glucose as its carbon source and 0.6% peptone plus 0.6% beef extract as nitrogen source after inoculating dosage 2mL suspension of conidia(1?107/mL).The optimal temperature and pH for enzyme activity is 50℃ and 6.0,respectively,while the activity can be enhanced by Tween-80 and inhibited by SDS.The enzyme activity is stable under 40℃ and in pH range of 5.5～6.5.

Objective To evaluate the relationship of peripheral blood Th17 and regulatory T (Treg) cells with disease activity in patients with systemic sclerosis (SSc).Methods This study recruited 21 patients with active SSc,24 patients with inactive SSc and 24 normal human controls with informed consent.Peripheral blood samples were obtained from these subjects.Flow cytometry was used to detect the percentages of Th17 and Treg cells in peripheral blood CD4+ cells,a fluorescence-based quantitative PCR to determine the levels of interleukin (IL)-17A,retinoid-related orphan receptor gamma t (RoRγt),forkhead box P3 (FoxP3) mRNA in peripheral blood mononuclear cells (PMBCs),and enzyme linked immunosorbent assay to measure the serum level of IL-17.Results Increased percentage of Th17 cells in peripheral blood CD4+ cells was observed in patients with active SSc compared with those with inactive SSc and normal human controls (2.34％ ± 1.19％vs.0.68％ ± 0.39％ and 0.57％ ± 0.49％,respectively,both P ＜ 0.05).No statistical difference was noted in the percentage of Treg cells in CD4+ cells or the mRNA expression levels of FoxP3 between the patients with active SSc,inactive SSc and normal human controls (all P ＞ 0.05).There was a significant increase in the mRNA expression of IL-17A,RoRγt in PBMCs and serum levels of IL-17 in patients with active SSc compared with patients with inactive SSc and normal human controls ( 11.73 ± 0.80 vs.9.77 ± 1.30 and 10.79 ± 0.74,respectively,both P ＜ 0.05; 18.48 ± 1.09 vs.15.89 ± 1.48 and 17.77 ± 1.64,respectively,both P ＜ 0.05; 53.60± 9.90 pg/ml vs.15.18 ± 3.24 pg/ml and 15.53 ± 4.12 pg/ml,respectively,both P ＜ 0.05).The percentage of Th17 cells in CD4+ cells and serum IL-17 levels were both positively correlated with disease activity in patients with active SSc (r =0.675,0.644,respectively,both P ＜ 0.05).Conclusions Th17 cells are highly proliferative in patients with active SSc,which may be closely correlated with the activity of SSc.

BACKGROUND: Peroxisome proliferation-activated receptor-gamma (PPARγ) is the member of nuclear receptor superfamily, and closely related with the formation of atherosclerosis.OBJECTIVE: To investigate the relationship between PPARγ C161→T gene polymorphism and coronary atherosclerotic heart disease (CAHD).DESIGN: Randomized controlled experiment SETTING: Department of Cardiology, Tonai Hospital of Huazhong University of Science and Technology; Department of Cardiology, Zhongnan Hospital of Wuhan University; Center for Human Genome Research,Huazhong University of Science and Technology; Department of Internal Medicine, Affiliated Hospital of Hubei University of Traditional Chinese Medicine; Institute of Geriatrics, Dongzhimen Hospital, Beijing University of Chinese Medicine PARTICIPANTS: Totally 203 CAHD patients aged (65±11) years, including 129 males and 74 females, were the inpatients and outpatients of Zhongnan Hospital of Wuhan University and Tonai Hospital of Huazhong University of Science and Technology from June 2002 to December 2005.And 156 cases of them were diagnosed by coronary arteriongraphy, among which 43 patients without coronary artery affection or with coronary stricture ＜ 50%, and 113 patients with coronary stricture ＞ 50 %. While 89 healthy physical examinees of Han race and mean (59±9) years old were enrolled as control group, including 56 males and 33 females. There was no blood relationship between controls and patients.METHODS: The experiment was conducted at Tongji Hospital of Huazhong University of Science and Technology from June 2002 to December 2005. PPARγ C161→T gene polymorphism was determined by polymerase chain reaction and restriction endonuclease fragment length polymorphisms. The radio-immunity technique, coronary angiography and clinical routine biochemical index were applied to analyze the genotypic frequency and allele frequency distributions as well as the relation between clinical data, biochemical index and different genotypes. The risk factors of CAHD were estimated in the patients of different genotypes.MAIN OUTCOME MEASURES: The genotypic frequency and allele frequency distributions, the relation between clinical data, biochemical index and different genotypes, along with the blood lipid, blood glucose, fasting insulin and body mass index (BMI).RESULTS: Totally 103 CAHD patients and 89 controls were involved in the result analysis of gene polymorphism and yielded different gene distribution frequencies.① In control group, "T" allele frequency was 0.213 and "C" allele frequency was 0.787, and in CAHD group, "T" allele frequency was 0.192 and "C" allele frequency was 0.808. There was no significant difference in the genotypic frequency and C, T allele frequencies between two groups (P ＞ 0.05).② The CC genotype was dominant in CAHD patients with coronary artery lesions, and showed significant differences from "T"allele carriers (CT+TT) (P ＜ 0.05). The CAHD risk in the "T" allele carries (OR: 0.56, 95% CI: 0.24-0.63) was much lower than that in the CC homozygote (OR: 1.92, 95% CI: 1.09-2.54).③ Apolipoprotein B in patients with CC genotype was obviously higher than that in patients with "T" allele (CT+TT) (P ＜ 0.05), and there was insignificant difference in the insulin resistance index (P ＞ 0.05).CONCLUSION: There is an important correlation between the substitution of PPARγ C161→T and CAHD, and "T" allele carriers demonstrate a lower risk of CAHD.

To observe the effects of total flavones of Metasequoia glyptostroboides Hu et Cheng (TFM) on volume-overload cardiac hypertrophy and the expression of c-Fos protein in rat. Methods　Volume-overload cardiac hypertrophy of rat was induced by aortocaval shunts. The rats were given ig TFM (400, 40 and 4 mg/kg/d). c-Fos protein in the ventricles were measured by immunocytochemical study. Results　TFM at the above dosage decreased heart weight and contents of RNA and protein in the myocardium, inhibited the expression of c-Fos protein in the ventricles. Conclusion　TFM can prevent volume-overload cardiac hypertrophy in rats. The inhibitory effects on the expression of c-Fos protein may be its mechanism in the molecular level.

Objective To investigate the interference and associated mechanism of hnman tissue kallikrein (HK) gene on renal interstitial fibrosis in rats with 5/6 nephrectomy. Methods Human kallikrein cDNA was packed in a recombinant adeno-associated virus(rAAV)-based plasmid vector. The rAAV-HK was produced by transfection in 293 cells. Twenty-four male Wistsr rats were divided into sham operation and operation groups. The rats with 5/6 nephrectomy were randomly divided into simple operation, control and experiment groups. The rats in experiment group received single dose rAAV-HK via the tail vein with 1×1011 pfu. Before nephrectomy and every month after surgery until the rats were sacrificed, the caudal arterial pressure was measured using tail cuff blood pressure determinator. Three months after HK gene delivery, the rats were sacrificed. The expression of HK in rats was assessed by RT-PCR, Western blot and enzyme-linked immunosorbent assay (ELISA). The pathological changes of renal interstitium were evaluated by Masson stainning, and the distribution of bradykinin B2 receptor (BKB2R) and angiotensin Ⅱ typel receptor (ATIR) was examined by immunohistochemistry. The expressions of BKB2R, AT1R, p-MAPK protein in renal tissue were detected by Western blot. Results Three months after HK gene delivery, the systolic blood pressure of experiment group was significantly decreased compared with the control group [(163±13) nun Hg vs (217±16) mm Hg, P<0.01](1 mm Hg=0.133 kPa). Compared with sham rats, the rats in simple operation group and control group had much more renal interstitial collagen deposition and more serious fibrosis performance, but renal interstitial collagen deposition and fibrosis were significantly ameliorated in the rats of experiment group. In addition, the tubulointerstitial injury index of HK transgenic rats was significantly lower than that of the rats in control group (1.33±0.73 vs 3.01±0.62, P<0.01). Up-regnlating expression of bradykinn B2 receptor protein and down-regulating expression of AT1 receptor and p-MAPK protein were found in renal tissues of experimental group after three months (P<0.05). Conclusion HK gene delivery significantly alleviates renal interstitial fibrosis in rats with 5/6 nephrectomy through regulating the expression of bradykinin B2 receptor, AT1 receptor and p-MAPK in renal tissue.