Adenomatous Polyposis Coli Protein ; genetics ; Chromatography, High Pressure Liquid ; methods ; Codon ; genetics ; DNA Mutational Analysis ; Fibromatosis, Aggressive ; genetics ; pathology ; Humans ; Mutation ; Polymerase Chain Reaction ; beta Catenin ; genetics

Objective To study the relationship between hepatocellular carcinoma and the interaction of polymorphisms in DNA repair gene XPD with environmental factors. Methods A hospital-based ease-control study on hepatoeellular carcinoma was conducted. All the hepatocellular carcinoma eases (n=300) were newly diagnosed and controls (n=312) were diagnosed with non-tumor cases. XPD genotype (Lys751 Gin and Asp312 Ash) from blood derived DNA was determined using TaqMan MGB Real-time PCR. Unconditional logistic regression was used to estimate the odds ratios (ORs) and 95% confidence intervals (CIs). Results For XPD condon 751 genotypes, there was no significant difference between frequencies of the AC or CC among patients and controls (P>0.05) (referent AA). The frequency of XPD312A allelic gene was higher in eases than that in controls and was associated with an increased risk (adjusted OR = 2.62,95% CI: 1.626-4.222) for hepatocellular carcinoma when compared with GG genotype. Interactions were found between infection of HBsAg and XPD312 (OR=7.348), as well as between smoking and non-wild type gene of XPD751 (0R=4.291) and XPD312 (OR=5.341). Conclusion DNA repair XPD312A allelic gene might increase the risk of Hepatocellular carcinoma. Interactions between HBsAg infection, smoking and XPD were observed in Hepatocellular carcinoma.

OBJECTIVETo summarize the pathogenic characteristics of bacterial infection and analyze the risk factors after orthotopic liver transplantation (OLT) in patients over 60 years of age.

METHODSA retrospective study of 69 patients that were over 60 years of age and underwent OLT was carried out. Descriptive statistics and risk factor analysis were performed with SPSS 11.0.

RESULTSThirty-eight patients developed bacterial infection (55.1%) after OLT, and thirty recipients suffered from mixed bacterial infection (79.0%). Multi-location infection was most commonly seen (68.4%). Nine patients died of bacterial infection. The primary pathogenic germs included enterococcus, methicillin-resistant coagulase negative staphylococcus, c maltophilia. The risk factors related to bacterial infection included preoperative malnutrition, long anhepatic phase, use of ventilator and duration of ICU stay.

CONCLUSIONSThe old patients that have undergone OLT are susceptible to bacterial infection. Bacterial infections are associated with high rate of mortality and multidrug resistance. Eliminating various risk factors can reduce the incidence of bacterial infection.

Aged ; Bacterial Infections ; etiology ; microbiology ; prevention & control ; Drug Resistance, Bacterial ; Female ; Humans ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Complications ; etiology ; microbiology ; prevention & control ; Retrospective Studies ; Risk Factors

OBJECTIVETo explore the role of abnormalities of chromosome 8, APC and beta-catenin genes in tumorigenesis of aggressive fibromatosis.

METHODSTrisomy 8 was detected by interphase fluorescence in situ hybridization (FISH). The APC gene and beta-catenin gene mutations were detected by denaturing high performance liquid chromatography (DHPLC) and direct sequence analysis after the PCR transition.

RESULTSThe rate of trisomy 8 in recurrent tumors (62.5%, 5/8) was significantly higher than that in the primary tumors (8.3%, 1/12). Somatic substitution of APC gene was found in 18 of 69 (26.1%) aggressive fibrometases. Somatic transition of beta-catenin gene was detected in 13 of 69 (18.8%) and mutation at codon 41 in exon 3 involving threonine residues implicated in the degradation of beta-catenin. The abnormal expression of beta-catenin had no significant correlation with the mutation of APC or beta-catenin gene. The group with positively expressed beta-catenin protein showed a significant higher c-myc protein expression than those without (P = 0.001). The Ki-67 index was extremely low in all the lesions. The apoptosis index (AI) of the groups with positively expressed c-myc and cyclin D1 showed significantly lower AI than those without.

CONCLUSIONTrisomy 8 may serve as a useful predictor of recurrence in aggressive fibromatosis. There are somatic mutations of the APC and beta-catenin genes in the aggressive fibromatosis, and there are abnormalities in the Wnt signaling pathway. These abnormalities may result in the aberrances of cell proliferation and apoptosis, which are likely to be import factors in the tumorigenesis.

Adenomatous Polyposis Coli Protein ; genetics ; metabolism ; Apoptosis ; Chromosomes, Human, Pair 8 ; Cyclin D1 ; metabolism ; Fibromatosis, Aggressive ; genetics ; metabolism ; pathology ; Genes, APC ; Humans ; Ki-67 Antigen ; metabolism ; Neoplasm Recurrence, Local ; Point Mutation ; Proto-Oncogene Proteins c-myc ; metabolism ; Signal Transduction ; Trisomy ; Wnt Proteins ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVETo study the clinicopathological and genetic features of desmoid-type fibromatosis, and to investigate the feasibility of detecting trisomy 8 in formalin fixed, paraffin embedded (FFPE) tissue by fluorescence in-situ hybridization (FISH).

METHODSA total of 96 cases were included in this study. All patients had clinical information. Histopathologic and immunohistochemical evaluations were available in 69 cases, and ultrastructural evaluation was done in 2 cases of desmoid-type fibromatosis. FFPE tissue sections were available in 20 tumors for the trisomy 8 detection by FISH.

RESULTSThere were 20 male and 76 female patients with ages ranging from 8 to 86 years (mean 35.3 years). Clinically, there were 44 extra-abdominal tumors, 28 abdominal wall tumors and 23 intra-abdominal lesions mostly involving the mesentery. Most cases presented with nodular or funicular masses with white firm cut surfaces, measuring 0.6 to 24.0 cm (mean 8.4 cm) in size. Histologically, desmoid-type fibromatoses showed longitudinal fascicles of spindle fibroblasts and myofibroblasts in a predominantly collagenous background. The tumor cells stained positive for vimentin, alpha-smooth muscle actin, desmin, and beta-catenin (47.8%, 33/69). Ultrastructurally, most tumor cells had features of fibroblasts, including rich endoplasmic reticulum and Golgi apparatus. Some tumor cells were myofibroblast-like cells exhibiting intercellular junctions, fibronexous junctions and stress fibers. Trisomy 8 was detected in 6 of 20 cases of desmoid-type fibromatosis including 5 of the 8 recurrent tumors but only one of the 12 primary tumors. The latter tumor also recurred three years later.

CONCLUSIONSDesmoid-type fibromatosis is an intermediate (locally aggressive) tumor that occurs predominantly in young females. The lesion consists of fibroblasts and myofibroblasts with the latter showing characteristic features including stress fibers and fibronexous junctions. Trisomy 8 can be detected in FFPE tissue by FISH, and its presence serves as a useful predictor of tumor recurrence and may define a subtype of desmoid-type fibromatosis with high recurrence rate.

Actins ; analysis ; Adult ; Aged ; Aged, 80 and over ; Child ; Chromosomes, Human, Pair 8 ; genetics ; Desmin ; analysis ; Feasibility Studies ; Female ; Fibromatosis, Abdominal ; genetics ; metabolism ; pathology ; Fibromatosis, Aggressive ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Male ; Mesentery ; Middle Aged ; Muscle, Smooth ; chemistry ; Neoplasm Recurrence, Local ; Peritoneal Neoplasms ; genetics ; metabolism ; pathology ; Trisomy ; Vimentin ; metabolism ; beta Catenin ; analysis

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; Female ; Fibromatosis, Aggressive ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged

OBJECTIVETo establish determination method of formic acid, lactic acid, acetic acid and succinic acid in dental plaque with high performance liquid chromatography (HPLC).

METHODSAfter the samples were centrifuged, 2 microL supernatant was transferred to a 1 mL centrifuge tube and diluted in water, then was determined with HPLC. The mixture of phosphate buffer and methanol (97:3) as mobile phase throughout the experiment. The determination of organic acid was performed on Phenomenex C18 column and at their maximum absorption wave.

RESULTSThe linear ranges of formic acid, lactic acid, acetic acid and succinic acid were 0.110-500, 0.049-500, 0.047-500, 0.084-500 microg/mL. The detection limits were 0.110, 0.049, 0.047, 0.084 microg/mL. The relative standard derivation were 9.5%, 7.9%, 4.3%, 4.2%. The average recoveries of samples were 82%-112%, 82%-102.5%, 90%-115%, 80%-110%.

CONCLUSIONThe method was simple, quick and adapt for analysis of organic acid in dental plaque.

Chromatography, High Pressure Liquid ; Dental Plaque ; Formates

OBJECTIVETo study the proteomic change in lymphocytes of rabbits with scald injury and Staphylococcus aureus (SA) invasion.

METHODSTwenty-four rabbits were divided into four groups as follows: control group, scald group, scald with SA invasion 2 hs group, and scald with SA invasion 6 hs group, according to random number table, with 6 rabbits in each group. Except for rabbits in control group (sham scald at 37 degrees C), rabbits in the other 3 groups were subjected to 30% TBSA full-thickness scald. Rabbits in SA invasion 2 and 6 hs groups were injected with 2 mL (1.0 x 10(8) CFU/mL) SA suspension, which was in the log growth phase, via auricle vein 18 hs and 22 hs after injury. Whole blood samples were collected from carotid artery of rabbits in 4 groups 24 hs after scald. Lymphocytes were isolated and its extracted proteins were analyzed by two-dimensional gel electrophoresis coupled with mass spectroscopy.

RESULTSAbout 1030 protein spots of lymphocytes were detected in each group. Compared with that of control group, 19 protein spots were found to be differentially expressed in the other 3 groups, and 11 spots (10 proteins) were identified. Expression levels of cofilin, cyclophilin A, ubiquitin, nucleoside diphosphate kinase, glutamate dehydrogenase and annexin 1 were down-regulated, but expression level of peroxiredoxin was up-regulated obviously.

CONCLUSIONSThere is obvious proteomic change in lymphocytes of scalded rabbits or of scalded rabbits invaded by SA, and it may relate to immune suppression and sepsis after injury.

Animals ; Burns ; metabolism ; microbiology ; Lymphocytes ; metabolism ; Male ; Proteome ; Rabbits ; Staphylococcal Infections ; etiology ; metabolism

OBJECTIVETo explore the mechanism and effect of sodium/hydrogen exchanger (Na(+)-H(+) exchanger, NHE), amiloride, on vessel stenosis.

METHODSThirty-two adult male New Zealand white rabbits were randomly divided into groups of amiloride intervention (IG, n = 12), balloon injury (BG, n = 10) and sham-operation (SG, n = 10). A 2.5 mm x 20 mm Foley's tube was used to injury left side iliac artery in the IG and BG groups, whereas a same Foley's tube was inserted into the vessel without any injuries in the SG group. Amiloride (5 mg.kg(-1).d(-1)) was intraperitoneally injected 3 days before balloon injuries and the same dosage normal saline was used in the same way in the BG group for 28 days after operation. The rabbits were killed and the iliac arteries were stained with Hematoxylin-Eosin, alpha-actin and Masson's trichrome to observe the morphologic changes in the vessel cava, neointima, media layer, and vascular smooth muscle cells (VSMCs) migration into the neointima and extracellular matrixes (ECMs).

RESULTSFour weeks after balloon injuries in rabbits, a cave narrow of the iliac artery and neointima were found and the media layer (VSM layer) was proliferated. The quantities of NHE-1 protein from artery smooth muscle in all the groups were 0.21 +/- 0.02, 0.25 +/- 0.04 and 0.11 +/- 0.03 (P < 0.01), respectively. The difference between the BG and SG groups was significant, which indicated that the NHE-1 proteins increased after balloon injury. The quantities of NHE-1 protein from the IG group were lower than those from the BG group. The cave areas were 0.91 mm(2) +/- 0.23 mm(2), 0.68 mm(2) +/- 0.19 mm(2) and 1.08 mm(2) +/- 0.17 mm(2) (P < 0.01), respectively. The intima areas were 0.27 mm(2) +/- 0.15 mm(2), 0.67 mm(2) +/- 0.24 mm(2) and 0.05 mm(2) +/- 0.03 mm(2), respectively (P < 0.01). The ratios of intima to media area were 1.21 +/- 0.24, 1.39 +/- 0.26 and 0.15 +/- 0.08 (P < 0.01), respectively. Amiloride increased vessel cave areas, but decreased intima areas and intima to media ratios in the IG group. In the IG group, alpha-actin positive areas in neointima was higher (16,328.31 microm(2) +/- 6220.27 microm(2)) than those in the SG group (4164.15 microm(2) +/- 1788.37 microm(2)) (P < 0.01). ECMs areas in neointima in the IG group were lower (8910.62 microm(2) +/- 7041.62 microm(2)) than those in the SG group (33,358.76 microm(2) +/- 7290.17 microm(2)) (P < 0.01).

CONCLUSIONSBalloon injuries of iliac artery in rabbits induce VSMCs proliferation, migration, narrowed cave and vessel stenosis. Amiloride, a NHE-1 inhibitor, may relieve this vessel stenosis.

Amiloride ; therapeutic use ; Angioplasty, Balloon ; Animals ; Blood Vessels ; injuries ; pathology ; Constriction, Pathologic ; Coronary Restenosis ; drug therapy ; metabolism ; pathology ; Iliac Artery ; injuries ; pathology ; Male ; Muscle, Smooth, Vascular ; drug effects ; metabolism ; Rabbits ; Sodium-Hydrogen Exchangers ; metabolism ; Stents

OBJECTIVEA reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of HIV-1.

METHODSRT-LAMP primers were designed according to conservative sequences of HIV-1 gag gene, and their sensitivity and specificity were evaluated by the established RT-LAMP protocol with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The performance of RT-LAMP on clinical samples was compared with real-time reverse transcription PCR(qRT-PCR).

RESULTSThe RT-LAMP assay showed a high specificity, and its detection limit was 1000 copies RNA per tube. The sensitivity and specificity of this method using 43 clinical samples were 94.6% and 100%, respectively,in comparison with those of qRT-PCR.

CONCLUSIONRT-LAMP assay using hydroxynaphthol blue dye does not need expensive instruments, and offer an alternative for the rapid detection of HIV-1 with the potential to be applied in field diagnosis.

HIV-1 ; isolation & purification ; Naphthalenesulfonates ; Nucleic Acid Amplification Techniques ; methods ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Reverse Transcription ; Sensitivity and Specificity