Aged, 80 and over ; Antibodies, Monoclonal ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Diagnosis, Differential ; Granuloma ; metabolism ; pathology ; Humans ; Keratins ; immunology ; Male ; Mucin-1 ; metabolism ; Skin Neoplasms ; metabolism ; pathology

AIMTo investigate the mechanisms of anti-cancer effect of resveratrol (Res), and the effects of Res in cell apoptosis. The role of Res playing in mitochondrial permeability transition pore (PTP) induction was studied.

METHODSMitochondria was prepared from the liver of Wistar rats. The effects of Res on oxygen consumption of isolated mitochondria from rat liver was measured with Clark-type electrode and resulted in respiration control rate (RCR). Mitochondrial swelling affected by Res was assessed spectrophotometrically, through the changes in absorbance at 540 nm. The PTP opening was learned from the results. Membrane potential of mitochondia was measured through fluorescence spectrophotometry.

RESULTSRes was shown to inhibit the respiration and decrease the RCR of mitochondria. Res can promote the PTP opening mediated by Ca2+. Res was shown to promote the increase of mitochondial membrane potential mediated by Ca2+ and loss of mitochondial membrane potential.

CONCLUSIONRes was shown to inhibit mitochondial respiration and induce PTP opening of mitochondria. These may be one of the pathways that Res showed anti-cancer action and induce cells apoptosis.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Calcium ; metabolism ; Female ; Ion Channels ; drug effects ; metabolism ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; drug effects ; physiology ; Mitochondrial Membrane Transport Proteins ; Mitochondrial Swelling ; drug effects ; Rats ; Rats, Wistar ; Stilbenes ; pharmacology

Objective To investigate the infections caused by Staphylococcus aureus carrying Panton-Valentine leukocidin(PVL)genes.Methods 26 isolates of Staphylococcus aureus carrying Panton- Valentine leukocidin(PVL)genes were determined by multiplex PCR.Multilocus sequence typing(MLST) was used to determine the STs of the isolates.The genotypes of SCCmec were also determined by another multiplex PCR in the isolates of methicillin-resistant Staphylococcus aureus(MRSA).Results Among 26 isolates,there were 6 isolates of ST88 MRSA,7 isolates of ST88 methicillin-susceptible Staphylococcus aureus (MSSA),5 isolates of ST239 MRSA,5 isolates of ST398 MRSA,1 isolate of ST25 MRSA,1 isolate of ST30 MRSA and 1 isolate of ST59 MRSA.20 isolates were hospital-acquired(HA)which mainly caused pulmonary infection and post-operative pyogenic infection.6 isolates were community-acquired(CA)which mainly caused soft tissue necrosis.Among 19 isolates of MRSA,ST88-SCCmec Ⅲ A,ST239-SCCmec Ⅲ,ST398- SCCmec Ⅳ and ST398-SCCmec Ⅲ were main types.26 isolates were isolated from 14 wards.ST88-SCCmec Ⅲ A-MRSA caused clone spread in maternity department in our hospital.Conclusion ST88,ST239 and ST 398 are main STs in Staphylococcus aureus carrying PVL in our hospital.The isolates not only cause nosocomial infections but also cause community infection.

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.

OBJECTIVETo find out how GATA-1 and GATA-2 behave in the bone marrow of patients with Monge's disease.

METHODSThe levels of mRNA in mononuclear cells (MNC) and proteins of GATA-1 and GATA-2 in the bone marrow of patients with Monge's disease and controls were determined by RT-PCR and immune cytolysis chemical method.

RESULTS(1) All patients and controls expressed GATA-1 mRNA (Monge's disease 1.033 +/- 0.146, Control 0.458 +/- 0.076) and GATA-2 mRNA (Monge's disease 0.451 +/- 0.073, Control 0.185 +/- 0.074). All patients expressed both GATA-1 (positive cell counts 77.3 +/- 33.3, positive score 135.4 +/- 75.4) and GATA-2 ( positive cell counts 29.4 +/- 11.4, positive score 48.4 +/- 19.7). All the controls expressed GATA-1 (positive cell counts 18.1 +/- 11.3, positive score 24.2 +/- 13.4) while 12 of 20 controls expressed GATA-2 ( positive cell counts 5.4 +/- 3.0, positive score 7.3 +/- 4.2). The expression of mRNA and proteins of GATA-1 and GATA-2 in Monge's disease were higher than in controls (P < 0.01). (2) There was a positive correlation between GATA-1 and Hb (P < 0.01), as did between mRNA and proteins of GATA-1 and GATA-2. (3) Both the proteins of GATA-1 and GATA-2 located only in the cytoplasm but not the nucleus.

CONCLUSIONSTwo of inherent genes, GATA-1 and GATA-2 which were expressed at higher levels in patients with Monge's disease than in controls might play significant roles in the pathogenesis of Monge's disease.

Adult ; Altitude Sickness ; metabolism ; GATA1 Transcription Factor ; metabolism ; GATA2 Transcription Factor ; metabolism ; Humans ; Male ; Polycythemia ; metabolism ; RNA, Messenger ; metabolism

OBJECTIVETo evaluate the impact of radiofrequency catheter ablation on left atrial (LA) size and function in patients with paroxysmal atrial fibrillation (PAF) and whether there is any difference between segmental pulmonary vein ostial isolation (SPVI) and circumferential pulmonary vein ablation (CPVA).

METHODSSixty-six patients with highly symptomatic atrial fibrillation were assigned to undergo either SPVI or CPVA. Transthorax echocardiography was performed before, 1 day, 1 months and 3 months after the procedure. LA dimension, LA area, late diastolic peak velocity of mitral valve inflow (A) and peak atrial systolic mitral annulus velocity (A') were recorded.

RESULTSOf 66 consecutive patients with symptomatic PAF, 30 patients underwent SPVI and 36 underwent CPVA. After a mean follow-up of (315 +/- 153) days, 21 patients (70%) after SPVI and 28 patients (75%) after CPVA were free of atrial tachyarrhythmia. As compared with the baseline, LA area decreased at 1-month after ablation in SPVI group and at 3-month in CPVA group. LA dimension decreased also in SPVI group, but did not in CPVA group. A velocity and A' velocity declined remarkably 1 day after CPVA, and restored 3 months later. The former went back to the level of baseline, and the latter exceeded it apparently. In SPVI group, A velocity increased at 1-month, and maintained in 3-month after ablation. A' velocity increased at 3-month after ablation. No reduction of A velocity or A' velocity was found after SPVI.

CONCLUSIONSThis study demonstrated a decrease in LA area and an improvement in LA systolic function 3 months after ablation for PAF. The LA damage by CPVA was more than that by SPVI, which was characterized by the reduction of LA function 1 day after procedure and the delayed improvement of LA size and functional parameters.

Adult ; Atrial Fibrillation ; diagnostic imaging ; physiopathology ; therapy ; Atrial Function, Left ; Catheter Ablation ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; Ultrasonography

Objective To investigate the genetic polymorphism of mec Ⅰ in the clinical isolates of methicillin-resistant Staphylococcus anreus(MRSA).Methods 40 isolates(MRSA)carrying mecA gene were selected randomly from the clinical isolates of Staphylococcus anreus from Jan,2005 to Aug,2006 in our hospital.The mec Ⅰ gene was detected by PCR followed with sequencing.Staphylococcal cassette chromosome mec(SCCmec)in MRSA were detected by multiplex-PCR.Agar dilution method was used for determining the MICs of oxacillin against MRSA.Results 35 of 40(87.5%)MRSA carried mec Ⅰ gene.All isolates carrying mec Ⅰ gene have mecI 202C→T substitution,which resulted in Gln at 68 aminophenol position replaced by stop condon.32 isolates carried single point mutation.3 isolates carried double-point mutation,including additonal A at 3 positon,A→C at 41 position and C→T at 142 position beside C→T at 202 position,respectively.Among 35 isolates carrying mec Ⅰ gene,there were 27 isolates of SCCmec Ⅲ, 7 isolates of SCCmec Ⅲ A and 1 isolate of SCCmec Ⅱ.Among 5 isolates with deletion of mec Ⅰ gene,there were 3 isolates of SCCmecⅣ,1 isolate of SCCmec Ⅰ and 1 isolate of non-known SCCmec tpye.The MICs of oxacillin were 256-512 ?g/ml,≥512 ?g/ml and 8-256 ?g/ml in 31 isolates with single point mutation at 202 position in mec Ⅰ gene,3 isolates with double-point mutation in mecI gene and 5 isolates with deletion of mec Ⅰ gene,respectively.1 isolate with single point mutation in mec Ⅰ gene had contrary result(MIC

OBJECTIVETo detect serve acute respiratory syndrome-associated coronavirus (SARS-CoV) and SARS-like-CoV in fruit bats captured in Guangzhou and its vicinity.

METHODSTotally 927 bats of 9 species (Cynopterus sphinx, Rousettus leschenaulti, Miniopterus schreibersi, Hipposideros pratti, Rhinolophusasinicus, Scotophilusakuhlii, Hipposideros Pomona, Rhinolophus affinis, and Rhinolophus pusillus) captured in Guangzhou and its vicinity from September 2004 to November 2005 were available for this investigation, from which 3,043 samples (813 throat swasb, 524 sera, 853 lung tissues and 853 colorectal tissue specimens) were obtained. SARS-Cov and SARS-like-CoV were detected in these specimens using diagnostic kit for novel coronavirus N protein (ELISA), SARS-CoV Virus RNA detection kit, fluorescence PCR, Genchip, RT-PCR and cell isolation culture methods.

RESULTS AND CONCLUSIONNo SARS-CoV and SARS-like-CoV were detected in the 3043 samples, indicating the current absence of SARS-CoV and SARS-like-CoV in the bats captured in Guangzhou and its vicinity.

Animals ; China ; epidemiology ; Chiroptera ; virology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nucleocapsid Proteins ; metabolism ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; metabolism ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology

BACKGROUNDTumor necrosis factor-alpha (TNF-α) is a pleiotropic proinflammatory cytokine and contributes to many kinds of cardiovascular diseases via its receptors (TNFR1/TNFR2). We hypothesize that TNF-α plays a role in the pathogenesis of chronic atrial fibrillation (AF).

METHODSSixty-seven consecutive patients who were scheduled to have cardiac surgery were enrolled into the study. Thirty-one patients with rheumatic heart disease (RHD) and AF were enrolled as study group (AF group). The sinus rhythm (SR) control groups consisted of 20 patients with RHD and 16 patients with coronary artery disease (CAD). Peripheral blood sample was collected before the operation. About 5 mm(3) left atrial tissue was disserted during the operation and was separated into three parts for Western blotting, real time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) analysis.

RESULTSCompared with the controls (RHD SR and CAD SR), the levels of TNF-α ((14.40 ± 5.45) pg/ml vs. (4.20 ± 3.19) pg/ml vs. (2.68 ± 2.20) pg/ml, P = 0.000) and its soluble receptor 1 (sTNFR1) ((1623.9 ± 558.6) pg/ml vs. (1222.3 ± 175.6) pg/ml vs. (1387.5 ± 362.2) pg/ml, P = 0.001) in plasma were higher in patients with AF. TNF-α level had positive correlation with the left atrial diameter (LAD) (r = 0.642, P = 0.000). Western blotting analysis showed that the protein levels of TNF-α (0.618 ± 0.236 vs. 0.234 ± 0.178 vs. 0.180 ± 0.103, P = 0.000) were higher in patients with AF. The RT-PCR analysis results demonstrated that the mRNA expression of TNF-α (0.103 ± 0.047 vs. 0.031 ± 0.027 vs. 0.023 ± 0.018, P = 0.000) increased in patients with AF. IHC analysis displayed that, comparing to the SR, the expression of TNF-α (0.125 ± 0.025 vs. 0.080 ± 0.027 vs. 0.070 ± 0.023, P = 0.000) increased in the AF group. The protein level and mRNA expression of TNF-α also had positive correlation with left atrium diameter (LAD) (r = 0.415, P = 0.000 and r = 0.499, P = 0.000).

CONCLUSIONSThe results revealed that TNF-α elevated in the plasma and left atrial tissue and had positive correlation with LAD in patients of chronic AF. TNF-α might involve in the pathogenesis of chronic AF.

Aged ; Atrial Fibrillation ; blood ; metabolism ; Blotting, Western ; Female ; Heart Atria ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; blood ; genetics ; metabolism

Zebrafish was selected as model animal, and glucocorticoid dexamethasone was used as a model compound to establish a rapid and high efficient osteopenia model. Zebrafish larvae at 4 days post fertilization (dpf) were exposed to a serial concentrations of dexamethasone solutions, and 0.5% DMSO was selected as the vehicle control group. All groups were incubated in 24-well plates (28.5 degrees C) until 9 dpf. In addition, effects of 10 micromol x L(-1) dexamethasone on preventing against osteopenia induced by etidronate disodium were also investigated. Zebrafish bones at 9 dpf were stained with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results showed that dexamethasone group at 2.5, 10 and 25 micromol x L(-1) can decrease the staining area and the staining optical density values of zebrafish head bones when compared with the vehicle control group (0.5% DMSO), which suggested that dexamethasone can significantly reduce the zebrafish mineralized bone and the bone mineral density. Results also showed that 15 and 30 microg x mL(-1) etidronate disodium can increase the mineralized matrix of zebrafish head bone and prevent against osteopenia induced by dexamethasone. In conclusion, the study indicated that zebrafish can be an idea osteopenia model induced by dexamethasone.
Animals ; Bone Density ; drug effects ; Bone Density Conservation Agents ; pharmacology ; therapeutic use ; Bone Diseases, Metabolic ; chemically induced ; prevention & control ; Calcification, Physiologic ; drug effects ; Dexamethasone ; toxicity ; Disease Models, Animal ; Etidronic Acid ; pharmacology ; therapeutic use ; Larva ; drug effects ; growth & development ; Zebrafish