Objective Genetic evaluation of relationship between rpoB gene mutation and rifampin-resistance of Mycobacterium tuberculosis (MTB).Methods This study was carded out with 81 clinical isolates and sensitivity test and genotypic analysis by PCR amplification and sequencing of rpoB gene were performed.Results Among these isolates,there were 27 rifampin-resistant and 54 sensitive strains.Of the 27 drug-resistant isolates,20 (74.1%) carried mutations on the amplified fragment of the rpoB gene with 10 mutation types at seven codons including 531 and 526,and two new mutation patterns were recognized.On the uther hand,one mutation (1.9%) appeared in 54 drug-sensitive strains.Conclusion The study shows geographical variation in the mutation types of rpoB gene in M.tuberculosis isolates from Guizhou,China.The result is valuable for the development of fast diagnostic methods.

Objective To explore the relationship of vascular endothelial growth factor(VEGF),IFN-? and IL-4 and effect of Bude-sonide on their in asthmatic rats.Methods Thirty-six male SD rats were randomly divided into 3 groups:asthma group,Budesonide-treatment group and control group.On the first day of the experiment and the 8th day,the rat models of the asthma group and Budesonid treatment group were allergized by the OVA/Al(OH)3 through intraperitoneal injection,respectively.And starting from the 15th day,they were challenged by the OVA through atomization for 2 weeks.Control group was allergized and challenged by NS atomization.Budesonid treatment group was interfered in Budesonide inhalation before suscitation in 0.5 h.After 12 h the same inhal done was again in Budesonide group.Twenty-four hours after the last challenge,the rats in 3 groups were sacrificed,and blood and bronchoalveolar lavage fluid(BALF) were collected.The concentrations of IL-4,IFN-? and VEGF in serum and BALF were measured by enzyme-linked immunosorbent assay.Results The concentrations of IL-4 in serum and BALF in asthma group and Budesonide treatment group were significantly increased than those in control group(P

OBJECTIVETo explore the effect of arsenic trioxide (As2 O3) on the level of VEGF, VEGFR and the activity of MMP-2, 9 in K562 cells.

METHODSThe inhibition ratio of K562 cell was detected by MTT assay, the level of VEGF by Enzyme-linked immunosorbent assay (ELISA), the expression ratio of VEGFR by flow cytometry (FCM), and the activity of MMP-2, 9 by gelatin zymography assay.

RESULTS(1) The IC50 of K562 cells was (2.12 +/- 0.11) micromol/L. Proliferation of K562 cells was significantly inhibited at the concentration of 0.4 - 6.4 micromol/L As2 O3 (P < 0.05). (2) The expression of VEGF was slightly up-regulated by 0.05 micromol/L As2 O3 (P > 0.05) and prominently inhibited by 0.4 micromol/L and 3.2 micromol/L As2 O3 (P < 0.05). As2 O3 had no influence on VEGFR. (3) The activity of MMP-2 and 9 was partly inhibited by 0.05 micromol/L As2 O3 incubated 72 hours and by 0.4, 3.2 micromol/L, As2 O3. With the increase of As2 O3 concentration and the incubation time, the inhibited effect on MMP-2 and 9 was enhanced.

CONCLUSIONSAs2 O3 may down-regulate the expression of VEGF and inhibit the activity of MMP-2 and 9.

Arsenicals ; pharmacology ; Cell Proliferation ; drug effects ; Humans ; K562 Cells ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Oxides ; pharmacology ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

The crude extracts of the fermentation broth from a marine sediment-derived actinomycete strain, Saccharothrix sp. 10-10, showed significant antibacterial activities against drug-resistant pathogens. A genome-mining PCR-based experiment targeting the genes encoding key enzymes involved in the biosynthesis of secondary metabolites indicated that the strain 10-10 showed the potential to produce tetracenomycin-like compounds. Further chemical investigation of the cultures of this strain led to the identification of two antibiotics, including a tetracenomycin (Tcm) analogs, Tcm X (1), and a tomaymycin derivative, oxotomaymycin (2). Their structures were identified by spectroscopic data analysis, including UV, 1D-NMR, 2D-NMR and MS spectra. Tcm X (1) showed moderate antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) pathogens, with the MIC values in the range of 32-64 microg x mL(-1). In addition, 1 also displayed significant cytotoxic activities against human cancer cell lines, including HL60 (leukemia), HepG2 (liver), and MCF-7 (breast) with the IC 50 values of 5.1, 9.7 and 18.0 micromol x L(-1), respectively. Guided by the PCR-based gene sequence analysis, Tcm X (1) and oxotomaymycin (2) were identified from the genus of Saccharothrix and their 13C NMR data were correctly assigned on the basis of 2D NMR spectroscopic data analysis for the first time.
Actinomycetales ; chemistry ; genetics ; Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Benzodiazepinones ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Data Mining ; methods ; Drug Resistance, Bacterial ; Enterococcus faecalis ; drug effects ; Fermentation ; Genomics ; Humans ; Inhibitory Concentration 50 ; Marine Biology ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Naphthacenes ; chemistry ; isolation & purification ; pharmacology ; Phylogeny ; Staphylococcus epidermidis ; drug effects

OBJECTIVETo detect gene mutations of children with glucose-6-phosphorate dehydrogenase (G-6-PD) deficiency and of carriers of G-6-PD deficiency gene with the technique of polymerase chain reaction and denatured gradient gel electrophoresis (PCR-DGGE), and to explore the value of the technique in the diagnosis of G-6-PD deficiency and G-6-PD deficiency gene carrying.

METHODScDNAs were harvested by reverse transcription method after RNAs had been extracted from peripheral blood of 43 children with G-6-PD deficiency and of their family members (36 lineages). Electrophoresis behaviors of the fragment from exons 11-12 of G-6-PD cDNA were detected with the technique of PCR-DGGE. Gene sequencing was then performed for the abnormal electrophoresis bands.

RESULTSAbnormal electrophoresis bands were found in the 1304-1520 fragment of G-6-PD cDNA in 33 out of 36 family lineages. The G-6-PD/6-PGD ratio was below 1.00 in 9 mothers of patients. Three of them had the G-6-PD/6-PGD ratio lower than 0.50. The PCR-DGGE bands were the same in the 3 mothers. Gene sequencing showed double heterozygote in the 3 mothers, but the maternal carriers of G-6-PD deficiency gene who had normal G-6-PD/6-PGD ratio showed mono-heterozygote in gene sequencing. Three mutational sites were found in the 1304-1520 fragment, i.e., C1311TG1376T and G1388A. The electrophoresis behaviors were different among the 3 gene mutational sites.

CONCLUSIONSPCR-DGGE is a sensitive and reliable technique in the screening of gene mutations. It is useful in the diagnosis of G-6-PD deficiency, especially in the diagnosis of female G-6-PD deficiency gene carrying.

Base Sequence ; Electrophoresis, Polyacrylamide Gel ; Female ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To detect miR-125a/b-5p expression in serum and urine of patients with preeclampsia,explore the role of miR-125a/b-5p in peripheral circulation on the pathogenesis of preeclampsia.Methods Samples of serum and urine of pa-tients with preeclampsia(n=20),and the normal pregnancy group(n=20)were collected and the expression of miR-125a/b-5p was detected by stem-loop-based real-time fluorescence quantitative reverse transcriptase polymerase chain reaction.Re-sults The expression of miR-125b-5p in the serum of patients with preeclampsia were significantly decreased compared with the normal pregnancy(Z=-2.272,P=0.023).There were no statistically differences of the expression of miR-125a-5p in serum between the two groups(Z=-0.622,P=0.547).The differences of urinary miR-125a/b-5p between patients with preeclampsia and normal pregnancy were not statistically significant(Z=-0.663,-0.189,P>0.05).Urinary miR-125b-5p was negatively correlated with diastolic blood pressure in preeclampsia group(r=-0.513,P=0.021).Serum miR-125b-5p was positively correlated with urea nitrogen in normal pregnancy group(r=0.472,P=0.036).Urinary miR-125a/b-5p were positively correlated with systolic blood pressure in normal pregnancy group(r=0.526,0.502,P=0.017,0.024). Conclusion The decrease of serum miR-125b-5p was associated with the pathogenesis of preeclampsia.

Objective To investigate the effects of parental support for the moderate-to-vigorous physical activity (MVPA) of their children and to promote physical activity for children and adolescents. Methods A total of 622 students were selected by stratified clustering sampling methods.Physical activity and parental support factors were assessed by self-report questionnaire survey.Logistic regression was used to analyze determinants of physical inactivity. Results Nearly 58.7% of the parents purchased sports equipment for their children and 57.6% of the parents accompanied their children to do exercise.The rate of parental explicit modeling was 22.8%.The students who received parental logistic support and explicit modeling did longer MVPA on weekends.The students with parental logistic support were prone to be in activity (OR=1.51, 95%CI:1.01-2.27). Conclusion Family-based interventions from parental logistic support and explicit modeling could promote middle-to-high-intensity activities for junior high school students.

OBJECTIVESTo introduce the method to monitor intrapelvic perfusion pressure during minimally invasive percutaneous nephrolithotomy (MPCNL), and to observe the effect of high pressure intrapelvic perfusion on recovery.

METHODSThe end of F5 ureteral catheter and aseptic transducer were connected by self-made connecter. During the operation, 46 renal calculi cases were monitored, and the early complication, such as fever, pain index, drop of hemoglobin, the stone-free rate and hospital stay were investigated. And the relationship between the variation of pressure and recovery was studied.

RESULTSIntrapelvic perfusion pressure ranged from 3 mm Hg (1 mm Hg = 0.133 kPa) to 50 mm Hg during the course of MPCNL. The definition of high pressure was the time of the pressure more than 30 mm Hg not less than 10 min. Postoperative fever rate, pain index, drop of hemoglobin and hospital stay in the high pressure were significantly higher than low pressure group (P< or =0.05). There was no difference of the stone-free rate in two groups.

CONCLUSIONSDuring the course of MPCNL intrapelvic perfusion pressure should be monitor immediately. It should be careful to maintain the time of pressure more than 30 mm Hg less than 10 min for stable postoperative recovery.

Adolescent ; Adult ; Female ; Humans ; Kidney Calculi ; physiopathology ; surgery ; Kidney Pelvis ; physiopathology ; Male ; Middle Aged ; Monitoring, Intraoperative ; Nephrostomy, Percutaneous ; methods ; Postoperative Complications ; physiopathology ; prevention & control ; Pressure ; Recovery of Function ; Retrospective Studies ; Young Adult

OBJECTIVETo study the clinical safety and effect on local recurrence in unresectable small hepatocellular carcinoma treated by radiofrequency ablation (RFA) with and without chemotherapy through a prospective randomized trial.

METHODSThirty-eight unresectable small hepatocellular carcinoma patients with diameter

RESULTSThere was no local recurrence of the tumor in the two groups 1 month after RFA treatment. The 6- and 12-month local recurrence rates were significantly lower in the combined group than that in RFA group alone (P < 0.01). There were no severe complications in the two groups, and nor was there any significant difference in liver function and WBC count.

CONCLUSIONRFA combined with systemic chemotherapy is safe, and it can reduce the local recurrence of unresectable small hepatocellular carcinoma
Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Hepatocellular ; drug therapy ; therapy ; Catheter Ablation ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Epirubicin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Liver Neoplasms ; drug therapy ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Prospective Studies ; Ultrasonography, Interventional

OBJECTIVETo detect the differential display code 3 mRNA (DD3 mRNA) in the urine sample of patients with prostate cancer and to evaluate its clinical significance.

METHODSDD3 mRNA in the urine collected from 48 patients with prostate cancer, 23 patients with benign prostate hyperplasia (BPH) and 9 healthy male volunteers was measured by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSDD3 products could not be detected in the urine samples of the BPH patients and healthy male volunteers, but could in 39/48 urine samples of the patients with prostate cancer. Significant difference was found between them (P < 0.01).

CONCLUSIONThe detection of DD3 mRNA in the urine promises to be a non-invasive, simple and sensitive method for the early diagnosis and post-treatment monitoring of prostate cancer.

Aged ; Aged, 80 and over ; Antigens, Neoplasm ; genetics ; urine ; Humans ; Male ; Middle Aged ; Prostatic Neoplasms ; diagnosis ; RNA, Messenger ; urine ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity