Differences in gene expressions were compared by cDNA microarray in skeletal muscle of type 2 diabetic rats and normal rats.In diabetic rats,157 genes were down-regulated and 100 genes up-regulated. Some of these genes were related to insulin resistance,glucose and lipid metabolism.

OBJECTIVE: To explore whether oxidized low-density lipoprotein (ox-LDL) can stimulate the cholesterol efflux in fully differentiated 3T3-L1 cells and the possible mechanism. METHODS: Fully differentiated 3T3-L1 cells were incubated in the medium containing various concentrations of ox-LDL ( 0 to 50 microg/mL) for 8 or 24 hours. 22(R)-Hydroxycholesterol (10 micromol/L) was exposed to preconditioned adipocytes with 25 microg/mL ox-LDL for 24 hours. Reverse transcription polymerase chain reaction (RT-PCR) was used to evaluate ATP binding cassette transporter A1 (ABCA1), scavenger receptor class B type I (SR-BI), and liver X receptor alpha (LXRalpha) mRNA expression. Cholesterol efflux mediated by apolipoprotein A-I (apoA-I) was determined using liquid scintillator. RESULTS: Low levels (12.5-25 microg/mL) of ox-LDL could increase cholesterol efflux via the enhancement of ABCA1 pathway and SR-BI expression, whereas the higher concentration (50 microg/mL) could not. In adipocytes preincubated with 25 microg/mL ox-LDL for 24 hours, 22(R)-hydroxycholesterol could increase ABCA1 and LXRalpha mRNA and apoA-I-mediated cholesterol efflux, but had no effect on the SR-BI mRNA expression. CONCLUSION Low levels of ox-LDL may enhance the LXRalpha-ABCA1-apoA-I pathway in adipocytes, up-regulate SR-BI mRNA expression, and then increase the cholesterol efflux. This new effect of ox-LDL will not only make contribution to cholesterol homeostasis in adipocytes, but also be potentially atheroprotective.
3T3-L1 Cells ; ATP Binding Cassette Transporter 1 ; metabolism ; Adipocytes ; drug effects ; metabolism ; Animals ; Cholesterol ; metabolism ; Lipid Metabolism ; Lipoproteins, LDL ; pharmacology ; Liver X Receptors ; Mice ; Orphan Nuclear Receptors ; metabolism ; Scavenger Receptors, Class B ; metabolism

Objective To evaluate the association between resting heart rate (HR) and all-cause death and coronary heart disease (CHD) events in the Chinese cohort. Methods Data were obtained from the PRC-USA Cooperative Study on Cardiovascular and Cardiopulmonary Epidemiology. Baseline screen surveys were conducted in 1983 and 1984 from people aged 35 to 59 years living in urban or rural areas of Beijing and Guangzhou. Follow-up visits were performed for end point events of all-cause death and first CHD events every two years till 2005. Resting HR was determined from 5 consecutive intervals between R waves on the 12-lead electrocardiogram. Results A total of 9856 (4805 males) people were included in the study and the mean follow up duration was 16. 2 years. There were 1523 deaths, including 200 CHD events during the follow up period. Mean resting HR was 67. 9 beat per minute (bpm) in men and 71. 6 bpm in women respectively which had a trend to increase with aging. Cox Proportional Hazards model indicated the relative risk of all-cause death increased constantly with the increase of HR percentile after control of age, fasting glucose, serum cholesterol, serum triglyceride, body mass index, systolic blood pressure and diastolic blood pressure. With HR 60-89 bpm as control group, the relative risk and 95% confidence interval in group HR <50 bpm,50-59 bpm,90-99 bpm and ≥100 bpm were 0. 76(0. 49-1. 17) ,0. 87 (0. 75-1. 02) , 1. 33 ( 1. 06-1. 68) ,1.48 ( 1. 03-2. 14) respectively. However there was no significant correlation between HR and CHD events in studied population. Conclusion The risk of total death increased significantly in people with HR≥90 bpm suggesting higher resting heart rate might be an independent risk factor for all-cause death in the Chinese population.

OBJECTIVEThis is a study on some ABO subgroup samples which show discordant results of serological and molecular blood typing, the aim is to clarify their true ABO type by means of nucleotide analysis on exons 6 and 7 of their ABO gene.

METHODSAbsorb-elution test and family investigation were conducted to study 7 samples which were involved in ABO grouping discrepancies. Duplex polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method was used to identify their ABO genotypes. PCR products of exons 6 and 7 were cloned and sequenced.

RESULTSAll the 7 ABO subgroup samples with the discordant results of serological and molecular blood typing were found to have the normal O gene. Four out of them were typed as ABsub by serology, they were all of the A*102/O genotype. Sequencing analysis found all their A gene having the nt467 (C-->T) and nt803 (G-->C) mutation by comparison with the A*101 allele, i.e. their real type should be CisAB/O. Three out of 7 were typed as AsubB by serology and as BO by genotype; and point mutation was detected in all of their B gene. One of them had the nt700 (C-->G) mutation, the other 2 unrelated individuals had the novel nt640 (A-->G) mutation in their B alleles.

CONCLUSIONThrough nucleotide analysis, 7 samples have been typed as AB subgroup in serology with the normal O gene, their real ABO type being CisAB in 4 cases and B(A) in 3 cases. At the same time, a kind of novel B (A)640 allele has been uncovered in this study.

ABO Blood-Group System ; genetics ; Asian Continental Ancestry Group ; genetics ; Blood Grouping and Crossmatching ; China ; Female ; Genotype ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

OBJECTIVETo construct lentivector carrying Tie2-Small interfering RNA (SiRNA), so as to study its influence on malignant melanoma cells.

METHODSRecombinant plasmid pSilencer 1.0-U6-Tie2-siRNA and plasmid pNL-EGFP were digested with XbaI, ligated a target lentiviral transfer plasmid of pNL-EGFP-U6-Tie2-I or pNL-EGFP-U6-Tie2-II, and then the electrophoresis clones was sequenced. Plasmids of pNL-EGFP-U6-Tie2-I and pNL-EGFP-U6-Tie2-II were constructed and combined with pVSVG and pHelper, respectively, to constitute lentiviral vector system of three plasmids. The Lentiviral vector system was transfected into 293T cell to produce pNL-EGFP-U6-Tie2- I and pNL-EGFP-U6-Tie2-II lentivirus. Then the supernatant was collected to determine the titer. Malignant melanoma cells were infected by both lentiviruses and identified by Realtime RT-PCR to assess inhibitory efficiency.

RESULTSThe recombinant lentiviral vectors of Tie2-RNAi were constructed successfully which were analyzed with restriction enzyme digestion and identified by sequencing. And the titer of lentiviral vector was 8.8 x 10(3)/ml, which was determined by 293T cell. The results of Realtime RT-PCR demonstrated that the lentiviral vectors of Tie2-RNAi could infect malignant melanoma cells and inhibit the expression of Tie2 genes in malignant melanoma cells (P<0.01). There was no significant difference in the expression level (P>0.05) between the two lentiviral vectors of Tie2-RNAi.

CONCLUSIONSLentivector carrying Tie2-SiRNA can be constructed successfully and inhibit the expression of Tie2 gene in vitro significantly. The study will supply the theory basis for the further research on the inhibition of tumor growth in vivo.

Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Melanoma ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, TIE-2 ; genetics ; Transfection

OBJECTIVETo develop a new nonisotopic detection method of enzyme-amplified time-resolved fluorescence (EATRF) or enzyme-amplified lanthanide luminescence (EALL) for nucleic acid hybridization assays, which can be applied extensively in clinical diagnosis.

METHODSThe method combines the high affinity of biotin-streptavidin system, amplification of enzyme, and inherent advantage of lanthanide chelate with the background elimination of time-resolved fluorescence detection. The conversion of 5-fluorosalicyl phosphate to 5-fluorosalicylic acid (5-FSA) by alkaline phosphatase. The salicylic acid product forms a luminescent ternary chelate with Tb3+ and EDTA.

RESULTSThe dynamic range of the standard curve of EATRFA for nucleic acid hybridization assay was very wide, the range was more than third order of magnitude. The detection sensitivity was about 10 pg of target sequence. When the known target sequence was 20, 10 and 2 ng, the ratio of measured amount to known amount was 110%, 90% and 115% respectively. The main experimental conditions, for example, the irradiating time of ultraviolet rays, the concentrations of biotinylated probe, AP-SA, 5-FSAP and Tb-EDTA and the methods of washing in the related steps, have been optimized. A new stable technology of fluorescence has been developted.

CONCLUSIONSEATRF detection for nucleic acid hybridization assays is a new sensitive simple method, which has a great prospect.

Alkaline Phosphatase ; analysis ; Blotting, Southern ; DNA ; genetics ; Fluoroimmunoassay ; methods ; Luminescent Measurements ; Metals, Rare Earth ; Nucleic Acid Hybridization ; methods ; Spectrometry, Fluorescence ; Substrate Specificity

OBJECTIVETo investigate the feasibility and effectiveness of transverse rectus abdominis musculocutaneous (TRAM) flap with partial preservation of abdominal rectus muscle based on the anatomic study in cadavers.

METHODS5 adult female cadavers which provided by department of anatomy of Fujian Medical University were dissected after injection with medical red latex from the starting point of the inferior epigastric artery and superior epigastric artery. The TRAM flap with partial preservation of lateral abdominal rectus muscle were dissected for breast reconstruction. The location, route, branches and anastomosis of inferior and superior epigastric arteries were observed. Based on the anatomic study, breast reconstruction were performed in 8 cases with muscle-sparing TRAM flaps.

RESULTSThe inferior epigastric artery arises from external iliac artery (9/10, 90%) or femoral artery (1/10, 10%) at the joint point between the internal third and lateral two third. There are extensive anastomoses between superior and inferior epigastric arteries above the umbilicus, mostly between the 2cm below the first tendinous intersection and umbilical level. From Sept. 2009 to Sept. 2010, 8 cases received breast reconstruction with muscle-sparing TRAM flap. The patients were followed up for 3 months to one year. Fibrosis happened in subcutaneous fat at flap IV zone in 2 cases, borderline necrosis and subcutaneous fat liquefaction occurred in some areas of flap IV zone in 2 cases, which healed after debridement. The other 4 cases healed with no complication. Except for unsatisfied shape in one case, good result achieved in 7 cases. There was no abdominal weakness, hemia or other complication.

CONCLUSIONSIt is an effective and safe method in breast reconstruction with muscle-sparing TRAM flap. It is practical with comparatively short operation time and less morbidity in donor site.

Adult ; Female ; Humans ; Mammaplasty ; methods ; Middle Aged ; Rectus Abdominis ; surgery ; Surgical Flaps

OBJECTIVETo investigate the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates from Chinese children in seven cities.

METHODA total of 134 MRSA isolates were collected from nine hospitals. Multilocus sequence typing and spa typing were analyzed by polymerase chain reaction (PCR), and staphylococcal chromosomal cassette mec (SCCmec) type was analyzed by multiplex PCR. The Panton-Valentine leukocidin (pvl) gene was also detected.

RESULTMost MRSA strains were isolated from pneumonia and skin and soft tissue infection (SSTIs) patients, accounting for 82.1%. Overall, 16 sequence types (STs) were obtained, and CC59 (51.7%) was found to be the most prevalent, which included ST 59 and ST 338, followed by ST239 (16.4%). SCCmec types II, III, IV, and V were also identified in the current study. SCCmec type IV was the most predominant type at 50.0%, followed by SCCmec type V at 23.9% and III at 23.9%. SCCmec subtypes IVa, IVc, and IVg were found among SCCmec type IV strains, whereas IVa was the main subtype at 77.6%. Twenty-six spa types were also identified, among which the predominant type was t437 (47.8%). The prevalence of pvl genes and the SCCmec type of strain was relevant, and the pvl gene positive rate was higher in SCCmec type IV and V-type strains than in SCCmec type II and III strains (58.6% vs. 14.3%, P < 0.05); there was a significant difference between them. In the strains isolated from pneumonia and SSTIs, ST59-MRSA-IVa(t437) was the predominant clone. There were five clones detected from the strains isolated from septicemia, with ST59-MRSA-IVa(t437) and ST59-MRSA-V(t437) as the main clones (57.1%). Various predominant clones existed in different regions. ST59-MRSA-IVa(t437) was the prevalent clone in the Guangzhou, Beijing, Chongqing, and Shenzhen areas, whereas ST239-MRSA-III(t037) was the prevalent clone in the Shanghai area. Fifty percent of the isolates from the Wenzhou area belonged to ST910-MRSA-V(t318), whereas three clinical strains isolated from the Shenyang region belonged to three different types.

CONCLUSIONThe results indicate that MRSA isolates from Chinese children are largely associated with the ST59-MRSA-IV(t437) and ST239-MRSA-III(t037) clones. These two may belong to community-acquired MRSA and hospital-acquired ones, respectively. Different prevalent clones were detected in different diseases and different regions. Therefore, there is a need to conduct further research on clinical isolates, which can guide the choice of antibiotic treatment and the examination of MRSA prevalence.

Adolescent ; Bacterial Typing Techniques ; Child ; Child, Preschool ; China ; epidemiology ; DNA, Bacterial ; genetics ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Methicillin-Resistant Staphylococcus aureus ; classification ; genetics ; isolation & purification ; Prevalence ; Staphylococcal Infections ; epidemiology ; microbiology

Objective To explore the mechanism of the intestinal barrier damage caused by Blastocystis hominis infections in rats. Methods Thirty SD rats were randomly divided into the control group, and the 1-, 3-, 6- and 9-week-infection groups, of 6 rats in each group. Rats in each infection group were orally infected with B. hominis trophozoites at a density of 2 × 10⁸ parasites per rat, and the control group was given an equal volume of phosphate buffered saline solution. The 7-hour urine samples were collected 1, 3, 6 and 9 weeks post-infection for the measurement of the intestinal permeability. Then, rats were sacrificed using the cervical dislocation method, and the cecum specimens were collected for the detection of the intestinal epithelial cell permeability. The expression of tight junction-related Occludin and Claudin - 1 genes and apoptosis-related Bcl - 2 and Bax genes was quantified in cecum epithelial cells using the real-time fluorescent quantitative PCR (qPCR) assay, and cell apoptosis was detected in the rat cecum using the TdT-mediated dUTP nick-end labeling (TUNEL) assay. Results The median urinary lactolose to mannitol ratios were 0.29, 0.72, 0.44, 0.46 and 0.38 in the control group, and the 1-, 3-, 6- and 9-week-infection groups, respectively, and the difference was statistically significant (H = 12.09, P < 0.05). B. hominis invasion and epithelial injury were observed in intestinal epithelial cells of rats infected with B. hominis, and transmission electron microscopy displayed the destruction of tight junctions between intestinal epithelial cells. The relative expression of Occludin, Claudin-1, Bcl-2 and Bax genes was 1.04, 0.62, 0.71, 0.68 and 0.96; 1.03, 0.61, 0.63, 0.76 and 0.86; 1.08, 0.70, 0.75, 0.74 and 1.03; and 1.00, 1.57, 1.33, 1.35 and 1.10 in the control group and the 1-, 3-, 6- and 9-week-infection groups, respectively, and all differences were statistically significant (F = 2.86, 2.85, 3.37 and 4.45, all P values < 0.05). The median number of positive staining cells were 1.00, 13.00, 9.00, 3.50 and 1.00 in rat cecum specimens in the control group, and the 1-, 3-, 6- and 9-week-infection groups, respectively, and the difference was statistically significant (H = 22.95, P < 0.01). Conclusion B. hominis infection may cause an increase in the rat intestinal permeability through triggering the apoptosis of intestinal epithelial cells to destroy the tight junction between intestinal epithelial cells, thereby destroying the intestinal barrier function.