Adult ; Aged ; Apoptosis ; Female ; Humans ; Male ; Middle Aged ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Pressure Ulcer ; metabolism ; pathology ; Skin ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Membrane ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Signal Transduction ; TCF Transcription Factors ; metabolism ; Transcription Factor 7-Like 2 Protein ; Wnt Proteins ; physiology ; beta Catenin ; metabolism

OBJECTIVETo study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.

METHODSE. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast.

RESULTSA total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79.

CONCLUSIONCTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.

Bacteremia ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; enzymology ; genetics ; isolation & purification ; Escherichia coli Infections ; microbiology ; Escherichia coli Proteins ; genetics ; Genotype ; Hospitalization ; statistics & numerical data ; Humans ; Liver Cirrhosis ; therapy ; Microbial Sensitivity Tests ; beta-Lactamases ; genetics ; metabolism

objecfive To know and compare the intelligence level of children born in different time periods in regions with iodine deficiency disorders(IDD)in Liaoning province.Methods All 7-14 year-old children from ten schools were chosen as the subjects respectively from six villages in each of the six counties and in regions with iodine deficiency,who were respectively born at the initialization of iodinated salt supplying period(1978-1980);non-iodinated salt supplying period(1981-1990);recovery of supplied iodized salt period(1991-1995);universal iodized salt period(1996-2000),respectively.Intelligence quotient(IQ)was measured by Combined Ravens Test in China(CRT-C)and Combined Ravens Test-the Rural,in China,2nd edition(CRT-RC2).Results IQ of children during the non-iodized salt period(91.9±14.3)was significantly lower than the initial supply of iodized salt period(95.8±14.6,q=8.60,P<0.01),recovery of supplied iodized salt period(99.7±14.7)was significantly higher than the initial supply of iodized salt period, non-iodized salt sales period(q = 9.53, 18.13, all P < 0.01 ),universal salt iodization( 104.3 ± 14.9) was significantly higher than the initial supply of iodized salt period, non-iodized salt sales period, recovery of supplied salt iodization(q = 20.00,28.00,10.46, all P < 0.01). Children's rate of mental retardation (IQ≤69) was higher in non-iodinated salt supplying period (6.7%, 88/1314 ) than the initial supply of iodized salt (4.4%, 21/471, χ2 = 3.85, P < 0.05), recovery of supplied iodized salt period(3.3%,48/1470) was significantly lower than non-iodinzed salt supplying period (χ2 = 15.37, P < 0.01), universal salt iodization period(2.7%, 36/1344) was lower than the initial supply of iodized salt period(χ2 = 4.41, P < 0.05) and non-iodinzed salt supplying period(χ2 = 26.34, P < 0.01 ). The IQ and intelligent retarded rates in children born during the initial years of iodinated salt supplying period were not different. The IQ of the children during ten years of non-iodized salt supplying period fluctuated in a "∪" curve, while the intelligent retardation rates in a "∩" curve.The children born during the period of recovery supplied iodized salt increased their IQ and lowered the retardation rates year after year. The IQ of the children in universal iodized salt period kept on increasing while intelligent retarded rates reduced to the lowest level. Conclusions The intelligence level of children born in regions with IDD during non-iodized salt supplying period is remarkably lower than that of the beginning years of iodinated salt supplying period. The intelligence level of children born after universal iodized salt period is remarkably higher than that of the initial iodinated salt supplying period and recovery of supplied iodized salt period, respectively.

OBJECTIVEBy establishing a model of straight-leg swaddle of newborn rats and observing the experimental animals'hips morphologically and pathologically, this study explored the changes of gross appearance of the acetabulum and the maturity of cartilage cells in the different regions of acetabular cartilage complex.

METHODSThe legs and hips were fixed by adhesive tape for 10 days in the position of hip extension and adduction in 31 newborn Wistar rats (experimental group). The other 31 newborn rats without legs and hips treatment were used as the control group. After 10 days raising in the same condition, all the rats were sacrificed. The gross appearance, histological observations and VEGF and type X collagen immunohistochemistry were used for examining the acetabulum changes.

RESULTSA straight leg swaddle model of newborn rats was established successfully. In the experimental group the acetabulum became shallow and small and surrounded by more soft tissues. There were 49 dislocated hips (49/54) in the experimental group and 2 hips dislocated (2/60) in the control group (p<0.01). Fake acetabulum appeared in the experimental group. In the control group, the shape of the acetabulum was normol, and no fake acetabulum was found. The safranin O-fast green staining showed that the orange-red cartilage in the experimental group was wider than the control group. Immunohistochemistry observations showed VEGF and type X collagen immunoreactivities in the hypertrophic layer of the acetabular cartilage complex in the experimental group were lower than those in the control group. The percentages of VEGF positive and type X collagen positive cells in the iliac hypertrophic layer of the acetabular articular cartilage were significantly higher than those in the ischiadic ramus and the pubic branch in the experimental group.

CONCLUSIONSVEGF and type X collagen immunoreactivities in acetabular cartilage cells decrease in a straight-leg swaddle model of newborn rats. This suggests that this position might lead to dysmaturity of the acetabular cartilage cells and affect the development of the acetabulum.

Acetabulum ; growth & development ; pathology ; Animals ; Animals, Newborn ; Bone Development ; Cartilage ; growth & development ; pathology ; Collagen Type X ; analysis ; Disease Models, Animal ; Female ; Hip Dislocation, Congenital ; metabolism ; pathology ; Immunohistochemistry ; Male ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; analysis

BACKGROUNDT cell factor-4 (TCF-4) plays an important role in development and carcinogenesis. Recently, the role of TCF-4 has been described in colon cancer and other cancers. However, whether TCF-4 plays a similar role in lung cancer is unknown. To answer this question, we studied the expression of TCF-4 protein and mRNA in non-small-cell lung cancer (NSCLC) and the relation of TCF-4 expression pattern to histological type and cell differentiation.

METHODSTissue samples from sixty cases of pathologically diagnosed NSCLC and eight normal tissue samples were obtained between September 2001 and March 2003. Immunohistochemistry was used to investigate the distribution of TCF-4 protein. The staining patterns of the tumors were divided into 4 categories: nuclear staining alone or nuclear staining greater than cytoplasmic staining; cytoplasmic staining or cytoplasmic staining greater than nuclear staining; equal nuclear and cytoplasmic staining; no nuclear staining or cytoplasmic staining. The integrated optical density (OD) values of all sections were analyzed by UIC MetaMorph image analysis software. The expression of TCF-4 mRNA was detected by one-step reverse transcription-polymerase chain reaction (RT-PCR). The integrated density values of the PCR products were analyzed semi-quantitatively.

RESULTSImmunohistochemistry showed that there was no expression of TCF-4 in normal tissue. However, TCF-4 was expressed in 86.7% (52/60) of NSCLC samples, mainly in the nuclei of tumor cells. Furthermore, there was a significant difference in TCF-4 localization patterns between squamous cell carcinomas and adenocarcinomas (P < 0.05). The integrated OD values of TCF-4 expression was significantly higher in tumors with moderate-poor cell differentiation than in well differentiated tumors (51.63 +/- 6.67 vs 46.13 +/- 12.31, P < 0.01). There was no TCF-4 mRNA expression in normal tissue. However, 63.9% (23/36) of carcinoma samples expressed TCF-4 mRNA. TCF-4 mRNA expression was significantly higher in tumors with moderate-poor cell differentiation than in well differentiated tumors (P < 0.05). There were no significant differences in mRNA expression in comparison with histological type.

CONCLUSIONSThe sub-cellular distribution of TCF-4 may correlate with NSCLC histological type. High expression of TCF-4 mRNA and protein may be associated with the degree of cell differentiation in NSCLC.

Carcinoma, Non-Small-Cell Lung ; chemistry ; Cytoskeletal Proteins ; metabolism ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; chemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; TCF Transcription Factors ; Trans-Activators ; metabolism ; Transcription Factor 7-Like 2 Protein ; Transcription Factors ; analysis ; genetics ; beta Catenin

OBJECTIVETo study the protective effect of the Mixture of Shengmai Powder and Danshen Decoction (, abbreviated as the Mixture) in the rat model with type 2 diabetic cardiomyopathy in the rat model with type 2 diabetic cardiomyopathy, abbreviated as the Mixture) in the rat model with type 2 diabetic cardiomyopathy (DCM).

METHODSForty-two SD rats with DCM model, established by the combination of insulin resistance by a high-fat diet with the damage of pancreatic islet β cells by intraperitoneal injection of high dose streptozotocin (50 mg/kg) once, were evaluated in the damage of the myocardium by electrocardiogram at the end of 12 weeks of grouping and intervention administration; the extent of damage in the myocardial subcellular structure was observed by electron microscopy; the content of myocardial collagen in the left cardiac ventricle was quantified by Masson staining test; the myocardial cell apoptosis was determined by TUNEL; the changes in the mRNA expression levels of thrombospodin-1 (TSP-1) and tribbles homolog 3 (TRB-3) by real-time quantitative PCR, the expression levels of myocardial TSP-1, tumor growth factorβ1 (TGF-β1), TRB-3, and chymase were detected by immunohistochemistry, and the changes in the expression levels of myocardial TSP-1, active-TGF-β1 (A-TGF-β1) and latent-TGF-β1 (L-TGF-β1) protein were tested by Western blotting.

RESULTSCompared with the control group, the myocardial tissue was less damaged, and the extent of damage in the myocardial subcellular structure was less; the collagen fiber content and the cell apoptosis were reduced; the expression levels of TSP-1mRNA and TRB-3 mRNA, the expression levels of myocardial TSP-1, TGF-β1, TRB-3, and chymase, as well as the average expression levels of the myocardial TSP-1, A-TGFβ1, and L-TGF-β1 protein were decreased in the Mixture group.

CONCLUSIONThe Mixture of Shengmai Powder and Danshen Decoction could inhibit the process of myocardial fibrosis in the rat myocardium of DCM through multiple pathways and significantly delay the genesis and progress of DCM in hyperglycemic rats.

Animals ; Cytoprotection ; drug effects ; Diabetes Mellitus, Experimental ; complications ; drug therapy ; Diabetic Cardiomyopathies ; pathology ; prevention & control ; Disease Models, Animal ; Drug Combinations ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Heart ; drug effects ; Male ; Myocardium ; cytology ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

OBJECTIVETo investigate the influence of pyrrolidine dithiocarbamate (PDTC) on the expression of transforming growth factor beta(1) (TGF-beta(1)), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor-1 of metalloproteinase (TIMP-1) in rats with pulmonary damage induced by paraquat (PQ).

METHODSFifty-four healthy male SD rats were randomly assigned into the control group (normal saline), the PQ-treatment groups (4 groups) and the PDTC treatment groups (4 groups). Except the rats in the control group, the rats in the PQ group were gavaged only with 40 mg/kg PQ, and PDTC group with 40 mg/kg PQ plus immediate injection 120 mg/kg PDTC (i.p). On the 3rd, the 7th, the 14th and 28th day after treatments, one group rats of each treatments were sacrificed and lung and blood samples were collected. The level of TGF-beta(1) protein in the plasma, the mRNA expression of TGF-beta(1), MMP-2 and TIMP-1 were evaluated using RT-PCR and real-time quantitative PCR, while pathological changes of lung were examined under optical microscope and electrical microscope.

RESULTSThe TGF-beta(1) protein, TGF-beta(1) and MMP-2 mRNA expression were increased significantly in the earlier stage and then decreased after PQ administration (P < 0.05 or P < 0.01), while the mRNA level of TIMP-1 was augmented continuously (P < 0.01) throughout the study compared to the control group. In comparison with the PQ group, in the PDTC treatment group, the TGF-beta(1) mRNA expression on the 3rd and the 14th day, 0.54 +/- 0.08 and 0.72 +/- 0.04 respectively, the MMP-2 mRNA expression on the 7th and 14th day, 1.62 +/- 0.50 and 1.97 +/- 0.34 respective-ly, and the TIMP-1 mRNA on the 7th and 21st day, 1.79 +/- 0.21 and 2.00 +/- 0.34 respectively, were significantly decreased (P < 0.05 or P < 0.01).

CONCLUSIONPDTC could attenuate paraquat-induced up-regulation of TGF-beta(1) and its mRNA expression, MMP-2 and TIMP-1 mRNA levels, which indicates that PDTC may exert its protective effects on paraquat-induced pulmonary damage by alleviating the earlier inflammation damage and adjust-ing the balance between MMPs and TIMPs. However, further studies are still warranted to investigate and clarify the underlying mechanisms involved in this complicated process.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Paraquat ; poisoning ; Pyrrolidines ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thiocarbamates ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

Objective To establish and optimize a sensitive and specific quantitative realtime polymerase chain reaction(PCR)method for detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA)in liver tissue. Methods Specific primers and probes were designed to detect HBV DNA(tDNA)and cccDNA. A series of plasmids(3.44 × 100-3.44 × 109 copies/μl)containing a full double-stranded copies of HBV genome(genotype C)were used to establish the standard curve of real-time PCR. Liver samples of 33 patients with HBV related hepatocellular carcinoma(HCC), 13 Chronic hepatitis B patients(CHB)and 10 non-HBV patients were collected to verify the sensitivity and specificity of the assay. A fraction of extracted DNA was digested with a Plasmid-Safe ATP-dependent Dnase(PSAD)for HBV cccDNA detection and the remaining was used for tDNA and β-globin detection. The amount(copies/cell)of HBV cccDNA and tDNA were measured by a real-time PCR, using β-globin housekeeping gene as a quantitation standard. Results The standard curves of real-time PCR with a linear range of 3.44 × 100 to 3.44 × 109 copies/μl were established for detecting HBV cccDNA and tDNA, and both of the lowest detection limits of HBV cccDNA and tDNA were 3.44 × 100 copies/μl. The lowest quantitation levels of HBV cccDNA in liver tissues tested in 33 HBV related HCC patients and 13 CHB patients were 0.003 copies/cell and 0.031copies/cell, respectively. HBV cccDNA and tDNA in liver tissue of 10 non-HBV patient appeared to be negative. The true positive rate was increasing through the digestion of HBV DNA by PSAD, and the analytic specificity of cccDNA detection improved by 7.24 × 102 times. Liver tissues of 2 patients were retested 5 times in the PCR for detecting cccDNA and the coefficience of variations on cycle threshold (Ct)were between 0.224%-0.609%. Conclusion A highly sensitive and specific quantitative real time PCR method for the detection of HBV cccDNA in liver tissue was established and could be used for clinical and epidemiological studies.

Objective:To investigate the association between MIC allele polymorphism and susceptibility to Ureaplasma urealyticum infection. Methods:PCR-SSP and PCR-SBT were used to analyze the gene polymorphism of every one. Results:Twelve allele genes of MICA and five MICA-STR and fourteen MICB were found in the participants,the frequency of MICA?010 and MICB?009N were higher in ureaplasma urealyticum infection patients than in healthy controls(MICA?010:OR=3. 85,95%CI:2. 12-6. 99, Pc<0. 05;MICB?009N:OR=3. 22,95%CI:1. 33-7. 80,Pc<0. 05);the frequencies of MICA-A5. 1 and MICB?00502 were lower in ureaplasma urealyticum infection patients than in healthy controls(MICA-A5. 1:OR=0. 61,95%CI:0. 40-0. 94,Pc<0. 05;MICB?00502:OR=0. 58,95%CI:0. 40-0. 83,Pc<0. 05),the differences were statistically significant;MICA?010/010,MICA?01201/01201 homozygote were higher in ureaplasma urealyticum infection patients than in healthy controls(MICA?010/010:OR=14. 84, 95%CI:1. 90-115. 9,Pc<0. 05:MICA?01201/01201:OR=10. 83,95%CI:1. 35-86. 79,Pc<0. 05),the differences were statistically significant. The distribution frequency of MICB?00502/00502 in patients with ureaplasma urealyticum was higher,but the difference was not statistically significant(Pc>0. 05). Conclusion:MIC allele gene polymorphism may be associated with ureaplasma urealyticum infection.