Objective To explore the expression and regulatory mechanism of hypoxia inducible factor-1?(HIF-1?)during fracture healing. Methods Mouse models of tibia fracture healing were established,and callus samples were collected 1,3,7,14,21 and 28 days after fracture.The development of callus and new bone formation were evaluated with roentgenology,Micro-CT and tetracycline double labeling method,and the expression of HIF-1?,vascular endothelial growth factor(VEGF),Runx2 and ALP in callus were detected with RT-PCR,Western blotting and immunohistochemistry.The relationship between HIF-1? and fracture healing was analysed. Results The expression of HIF-1? was detected in cells in the fracture sites as well as in evolved osteoblasts,chondrocytes and osteocytes in early callus under hypoxia.The highest expression rate of HIF-1? achieved on the 7th day after fracture,lasted for about 7 days,then decreased gradually,and returned to intact level on the 28th day after fracture.The expression tendency of VEGF resembled that of HIF-1?.Bone formation activity was more active in early callus,and the callus volume peaked on the 14th day after fracture and decreased gradually.The mineralization of callus mainly took place in the late healing period(14th to 28th day after fracture).Conclusion Cells involved in fracture healing are hypoxia-responsive cells,which express HIF-1?.HIF-1? can regulate cell state and function,and can promote angiogenesis so as to play a crucial role in fracture healing.

Adolescent ; Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Humans ; Influenza A virus ; immunology ; Male ; Seroepidemiologic Studies ; Students, Medical ; Vietnam ; epidemiology ; Young Adult

OBJECTIVETo investigate the expression of long non-coding RNA HOTAIR in the plasma of breast cancer patients and its value in the diagnosis of breast cancer.

METHODSHOTAIR levels were measured in 24 tumor tissues and 70 plasma samples from breast cancer patients using quantitative real-time PCR. The correlations of plasma HOTAIR level with the clinicopathological features of the patients were analyzed. A multivariate logistic regression model was established to analyze the value of plasma HOTAIR in comparison with plasma CA153 and CEA levels for breast cancer diagnosis. We further detected HOTAIR levels in the plasma and breast cancer tissues of 24 patients before and after operation and investigated their correlation.

RESULTSBreast cancer patients had increased expressions of HOTAIR in the tumor tissues and plasma, and plasma HOTAIR level was significantly correlated with estrogen receptor (ER) level (P=0.004) and lymph node metastasis (P=0.010). Receiver operating characteristic (ROC) curve and the multivariable logistic regression model showed that the area under ROC curve (AUC) of plasma HOTAIR was 0.82 (P<0.001) for breast cancer diagnosis with a diagnostic sensitivity and a specificity of 73.3% and 93.3%, respectively. The diagnostic power and specificity of plasma HOTAIR was much higher than those of CA153 (AUC=0.66, P=0.030) and CEA (AUC=0.52, P=0.001), and the combination of the 3 markers further enhanced the diagnostic power (AUC=0.84) and specificity (96.7%). Plasma HOTAIR level was significantly reduced in the patients after the operation (P<0.0001) and showed a moderate correlation with its expression in tumor tissues (r=0.62, P<0.0001).

CONCLUSIONPlasma HOTAIR may serve as a potential biomarker for breast cancer diagnosis.

Biomarkers, Tumor ; blood ; Breast Neoplasms ; blood ; diagnosis ; Carcinoembryonic Antigen ; blood ; Female ; Humans ; Logistic Models ; Lymphatic Metastasis ; Mucin-1 ; blood ; Prognosis ; RNA, Long Noncoding ; blood ; ROC Curve ; Real-Time Polymerase Chain Reaction ; Receptors, Estrogen ; metabolism ; Sensitivity and Specificity

China ; Genes, Bacterial ; Sequence Analysis ; Streptococcus suis ; classification ; genetics ; isolation & purification ; Virulence Factors ; genetics

OBJECTIVETo investigate the prevalence of three lamivudine-resistant HBV mutants in lamivudine-treated Chinese patients.

METHODSUsing three pairs of of HBV polymerase gene B and C domain fragments were amplified by PCR. The PCR products were then digested by restriction enzyme Nde I and Nla III. The digested products were analyzed by electrophoresis. With this method, the prevalence of the three lamivudine-resistant mutants in lamivudine-treated Chinese patients was investigated.

RESULTSAfter Nde I digestion of p24 and p29 amplified product, HBV wild type could be easily separated from YMDD mutant. At the same time, YIDD could be separated from YVDD mutant after Nla III digestion of p24 and p29 amplified product. By this method, the authors found that these eleven patients were infected with lamivudine-resistant mutants. Six of them were infected with M5501 mutant; five were infected with M550V mutant (one of them had both M550V and L526M mutations).

CONCLUSIONThe method of the present study was demonstrated to be an easy way to detect HBV lamivudine-resistant mutants and can be applied to clinical monitoring of lamivudine resistance.

Adolescent ; Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA Mutational Analysis ; DNA Primers ; genetics ; DNA, Viral ; analysis ; Drug Resistance, Viral ; genetics ; Female ; Hepatitis B ; drug therapy ; virology ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Point Mutation ; Polymerase Chain Reaction

Objective To develop a rapid, sensitive and specific assay method, based on multiplex real time PCR for identifying Vibrio cholerae and distinguishing Vibrio cholerae O139 serotype from Vibrio cholerae. Methods Cholera toxin A subunit gene (ctxA) and glycosyltransferase gene (LPSgt) were chosen as targets according to Vibrio cholerae and Vibrio cholerae O139 serotype,and then the primers and TaqMan-MGB probe were designed. The 5'end of probes was labeled with FAM and VIC fluoresceins respectively while the 3' end of probes was labeled with MGB. The PCR reaction was optimized systematically. Then the specificity, sensitivity and reproducibility of multiplex real time PCR were estimated. Finally, multiplex real time PCR was applied to detect the clinical specimens. Results Vibrio cholerae was identified by multiplex real time PCR accurately and quickly,which could distinguish Vibrio cholerae O139 serotype from Vibrio cholerae. Vibrio cholerae was identified positive for primer pairs and probes from ctxA gene, and Vibrio cholerae O139 serotype tested was positive for LPSgt gene. Meanwhile, none of other bacteria was identified. Sensitivity of the test was 2 × 102 cfu/ml. When this assay was applied directly to identify 45 clinical specimens, results showed that 10 were positive to Vibrio cholerae, in which 4 clinical specimens were positive to Vibrio cholerae O139 serotype. All the results were the same to the one that had been obtained from the conventional assays. Conclusion Our rsults showed that the multiplex real time PCR was a reliable,accurate and feasible method for identifying Vibrio cholerae that carrying toxin gene and distinguishing Vibrio cholerae O139 serotype from Vibrio cholerae. This method could be applied to the cholera surveillance, prevention and control system for identifying and distinguishing Vibrio cholerae in the labs.

Objective To establish a real time PCR assay with a novel fluorescence quencher for identification of mutation of clarithromycin -resistance gene of Helicobacter pylori.Methods Two mutations of 23S rDNA gene in Helicobacter pylori,No.2142 and 2143,were chosen as targets for detection,and then the primers and the probe with a novel fluorescence quencher were designed.The genome DNA of Helicobacter pylori was extracted,and then detected by real time PCR reported here.Meanwhile,the specificity,reproducibility and sensitivity of the assay were evaluated.Finally,the real time PCR described here,the real time PCR based on TaqMan,and a sequencing assay were applied to detect 55 Helicobacter pylori strains isolated from clinical specimens,respectively.The results from three assays were compared with each other in order to further evaluate the applicability of this assay in clinic.Results It indicated that the mutation points related to clarithromycin -resistance,A2142G and A2143G,were identified by real time PCR with a novel fluorescence quencher rapidly and accurately.Moreover the coefficient of variation was less than 5%.The limit of detection was 100 copies/reaction.While this assay was applied directly to detect 55 Helicobacter pylori strains,the results were in accordance with those obtained from a TaqMan real time PCR and a sequencing assay,respectively.Conclusion The real time PCR described here was a simple,reliable and accurate approach and substituted for the TaqMan real time PCR for identification of two mutation points of clarithromycin -resistance,A2142G and A2143G in Helicobacter pylori.Thus,a novel tool for diagnosis of gene mutation was provided and the results might be regarded as a substantial evidence for clinical individual therapy.

OBJECTIVETo compare the image quality and radiation dose of prospectively electrocardiogram (ECG) -triggered spiral and sequential acquisition for coronary computed tomographic angiography by dual-source computed tomography.

METHODSSixty patients with suspected or known coronary artery disease were randomly divided into two groups. Group A underwent prospective ECG-triggering spiral scan and Group B underwent prospective ECG-triggering sequential scan. Both the image quality and radiation dose of the two groups were compared.

RESULTSThere was no significant difference in age and body mass index of the two groups. The average image quality score was 1.12 ± 0.38 in group A and 1.14 ± 0.38 in group B (Z=-0.291,P=0.771) . The rates of diagnostic coronary segments for two groups were 98.87% and 99.56% respectively (X2=0.59,P=0.443) . The mean radiation dose of group A was significantly lower than that of group B [ (1.31 ± 0.30) mSv vs. (3.36 ± 0.93) mSv; t=11.47, P=0.000] .

CONCLUSIONCompared with the prospective ECG-triggered sequential acquisition, the prospective ECG - triggered spiral scan for coronary computed tomographic angiography can remarkably reduce radiation dose without impairing image quality in patients with a low and stable heart rate (≤ 70 bpm) .

Aged ; Coronary Angiography ; methods ; Coronary Artery Disease ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Radiation Dosage ; Tomography, Spiral Computed ; methods ; Tomography, X-Ray Computed ; methods

Objective To investigate the clinical effect of compositive chest physiotherapy for the prevention of atelectasis after thoracotomy in the elderly patients.Methods Sixty-eight elderly patients after thoracotomy were randomly divided into two groups, the experimental group adopted compositive chest physiotherapy(n = 35), the control group were given the traditional chest physiotherapy(n = 33).The factors considered included the action abide by doctor,the effect of expectoration, the improvement difference of PaO2 ,PaCO2 , SaO2 by blood gas analysis, the incidence of atelectasis.Results There were significant differences in the action abide by doctor and the effect of expectoration. After intervention, the improvement difference of PaO2 ,PaCO2 and SaO2 were significant higher than that of the control group(P <0.05); The rates of atelectasis were 5.71% (2 cases) and 27.27% (9 cases) in experimental group and control group respectively (P < 0.05).Conclusions Compositive chest physiotherapy is superior to traditional physiotherapy for elderly patients in pedoperation, which can clear sputum more effectively, improve blood oxygen partial pressure and lung expansion, also can prevent pest-operative atelectasis more effectively in the elderly patients.

OBJECTIVETo investigate the distribution of beta-2-AR +46 A-->G variant in Kazakans of Xinjiang and the relationship of the variant with low density lipoprotein-cholesterol (LDL-C) level in this population.

METHODSThe genotypes of beta-2-AR gene Arg16/Gly variant were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in 506 Kazakans with age from 30 to 69, and its distribution and relationship to LDL-C level were investigated.

RESULTS(1) The frequencies of genotypes AA, AG, GG and alleles A, G of beta-2-AR +46 variant in this population were 0.310, 0.455, 0.235 and 0.538, 0.462 respectively, which were accorded with Hardy-Weinberg equilibrium. (2) The Gly16/Gly genotype had highest LDL-C level in the three genotypes, and were significantly higher than Arg16/Gly genotype (P < 0.05). (3) Comparing the effect of beta-2-AR gene +46 variant on serum lipid in males with females, we found that females with Gly16/Gly genotype had the highest level of serum LDL-C.

CONCLUSIONSThese data show that Gly16/Gly genotype of beta-2-AR gene +46 A-->G variant is associated with higher level of serum LDL-C in this population, especially in female.

Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; genetics ; Cholesterol, LDL ; blood ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Receptors, Adrenergic, beta-2 ; genetics