Objective To explore the significance of sample cultivation and drug resistance of the patients in PICU with severe lower respiratory tract specimens.Methods 1 107 pathogenic bacteria of lower respiratory tract samples collected were analyzed and the drug resistance was kept watch on.Results A total of 1107 samples were cultured into 948 strains of pathogens,with the positive rate of 85.64%,and 616 strains of Gram negative bacteria accounting for 64.98%,of which 145 strains of Pseudomonas aeruginosa(15.30%),Klebsiella pneumoniae 135(14.24%),Acinetobacter spp 131(13.82%),large intestine Egypt Greek bacteria 89(9.4%);Gram positive cocci 129(13.61%),the main pathogen was staphylococcus aureus and Fungi 203(21.41%),the main pathogen was candida albicans.Carbapenems was the most active antibiotics for gram negative bacill,and vancomycin for gram positive cocci.Conclusion Infectious bacteria have high selectivity to the sensitive antibiotics,and doctors should carefully use antibiotic according to the susceptibility test results in order to reduce hospital infection and the emergence and spread of resistant strains.

Objective:To investigate effects of miR-503 on cisplatin sensitivity in BEL-7402 cells by targeting of bcl-2.Methods:MiR-503 and bcl-2 mRNA expression levels in hepatocellular carcinoma cells were measured by real-time quantitative (qRT)-PCR;Bcl-protein level was detected by Western blot;miR-503 mimics were transiently transfected to the BEL-7402 cells by liposome transfection;potential target genes of miR-503 were predicted by Bioinformatics software;miR-503 potential targets were validated by dual luciferase activity;and the cell viability was measured by MTT assay.Results:MiR-503 level was down-regulated and Bcl-2 protein expression level was up-regulated in BEL-7402 cells compared with HL-7702 cells.MiR-503 could interact with bcl-2 and inhibit its expression.Cell vitality with miR-503 transfection was significantly reduced compared to that in the negative control.Conclusion:MiR-503 may enhance the sensitivity of BEL-7402 cells to cisplatin and inhibit the cell proliferation by targeting bcl-2.

OBJECTIVE CYP2D is one of the most abundant subfamily of CYPs in the brain, especially in the cerebellum. Brain CYP2D is responsible for the metabolism of endogenous neurotransmitters such as tyramine and serotonin. Our previous studies have shown brain CYP2D can be regulated by exogenous and endogenous substances with tissue- specificity. The purpose of this study is to examine the effects of cerebral CYP2D on the mice behavior and the regulatory mechanism of brain CYP2D by growth hormone. METHODS Mice received the stereotaxic injection with CYP2D inhibitor quinine in deep cerebellar nuclei of cerebellum. The animals were tested with rotarod apparatus, balance beam, water maze, elevated plus maze and open field. The changes in CYP2D22, PPARαand PPARγ in brain regions and liver were assayed in male growth hormone receptor knockout mice, SH-SY5Y cells and HepG2 cells. RESULTS The inhibition of cerebellum CYP2D significantly affected the spatial learning and exploring ability of mice. Compared with WT mice, CYP2D expression was lower in brain regions from GHR(-/- ) male mice; however, hepatic CYP2D level was similar. Pulsatile GH decreased PPARα mRNA level, and increased mRNA levels of CYP2D6 and PPARα in SH- SY5Y cells. In HepG2 cells, pulsatile GH resulted in decreases in PPARα and PPARγ mRNA levels, but not CYP2D6. PPARα inhibitor induced CYP2D6 mRNA and protein by 1.32-fold and 1.43-fold in SH-SY5Y cells. PPARγ inhibitor decreased CYP2D6 mRNA and protein by 74.76% and 40.93%. PPARα agonist decreased the level of CYP2D22 mRNA in liver and cerebellum, while PPARγ agonist rosiglitazone resulted in diametrically increases. The luciferase assay showed that PPARγ actived the CYP2D6 gene promoter while PPARα inhibited its function. Pulsatile GH declined the binding of PPARα with CYP2D6 promoter by 40%, promoted the binding of PPARγ with CYP2D6 promoter by approximate 60%. The levels of brain and liver PPARα expression in male GHR(-/- ) mice is obviously higher than those in WT mice. The level of PPARγ in male GHR(-/- ) mice was decreased in the frontal cortex and hippocampus, while remained stable in the cerebellum and striatum; meanwhile, PPARγ was increased in the liver. CONCLUSION Brain CYP2D may be involved in learning and memory functions of central system. Masculine GH secretion altered the PPARs expression and the binding of PPARs to CYP2D promoter, leading to the elevated brain CYP2D in a tissue- specific manner. Growth hormone may specifically alter the metabolic and synthetic of important endogenous substances in the central nervous system (such as serotonin) through the specific regulation of brain CYP2D expression.

Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
Genetic Engineering ; Macrolides ; metabolism ; Point Mutation ; Ribosomal Proteins ; genetics ; Ribosomes ; metabolism ; Saccharopolyspora ; metabolism

ObjectiveTo estimate the clinical efficacy and immunomodulatory effect of polysaccharidenucleic acid fraction of bacillus calmette guerin(BCG-PSN) combined with a traditional Chinese medicine in patients with vitiligo.MethodsThis study recruited 99 patients with vitiligo aged from 13 to 65(35,6 ± 5.8) years.The patients were classified into 3 groups to be treated with BCG-PSN and a traditional Chinese medicine (Baidianfeng granules) alone or in combination.BCG-PSN was intramuscularly injected at a dose of 2 ml every other day and baidianfeng granules were given orally thrice a day,for 3 months.Peripheral blood samples were obtained from the patients at the baseline and after the end of treatment and from 30 healthy controls.Flow cytometry and enzyme linked immunosorbent assay(ELISA) were performed to detect T cell subsets and expression level of Fas and Fas ligand(FasL),respectively.Data were analyzed by t test and chi-square test.Results The response rate was significantly higher in patients treated with BCG-PSN combined with Baidianfeng granules than in those with BCG-PSN alone(82.86％ vs.40.63％,P ＜ 0.01 ).Before the treatment,patients showed a lower percentage of CD3+ cells,CD3+CD4+ cells and CD3+CD8+ cells in peripheral blood(all P ＜0.01 ),weaker expression of Fas (P ＜ 0.01 ),but a higher CD4/CD8 ratio (P ＜ 0.01 ) compared with the controls.The treatment with BCG-PSN and Baidianfeng granules alone or in combinatiou all induced an increase in the percentage of CD3+ cells,CD3+CD4+ cells and CD3+CD8+ cells in peripheral blood(P ＜ 0.05) and in the expression of Fas(P ＜ 0.01),but a decrease in CD4/CD8 ratio(P ＜ 0.05).ConclusionsBCG-PSN may induce the normal apoptosis in lymphocytes via reversing the abnormality in the expression of Fas/FasL by peripheral blood lymphocytes,and the effect of BCG-PSN may be enhanced by a traditional Chinese medicine,Baidianfeng granules.

OBJECTIVETo investigate the role of mitochondrial calcium uniporter in cardioprotection elicited by ischemic postconditioning (Postcond).

METHODSMale Sprague-Dawley rats were used for Langendorff isolated heart perfusion. The hearts subjected to global ischemia for 30 min followed by 120 min of reperfusion. Left ventricular developed pressure (LVDP), maximal rise/fall rate of left ventricular pressure (± dP/dtmax) were measured. The level of lactate dehydrogenase (LDH) in the coronary effluent was measured spectrophotometrically, the content of formazan of myocardium was also measured at the end of reperfusion.

RESULTCompared to I/R group, Postcond had an significant increase in the mechanical function of the left ventricle, with LDH release reduced and the content of formazan increased. Spermine, the opener of mitochondrial calcium uniporter, deteriorated the mechanical function of left ventricle and decreased the formazan content, and increased LDH release. Ruthenium red, the inhibitor of mitochondrial calcium uniporter, increased the mechanical function of the left ventricle, decreased the LDH release, but the content of formazan was not increased.

CONCLUSIONThe inhibition of mitochondrial calcium uniporter is involved in the mechanisms of ischemic postconditioning.

Animals ; Calcium Channels ; metabolism ; physiology ; Disease Models, Animal ; Formazans ; analysis ; Heart ; physiopathology ; Ischemic Postconditioning ; L-Lactate Dehydrogenase ; analysis ; Male ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; prevention & control ; Rats ; Rats, Sprague-Dawley

Objective: To evaluate the impact of rehabilitation on exercise capacity, cardiopulmonary function and quality of life (QoL) in patients with chronic heart failure (CHF). Methods: A total of 96 CHF patients with NYHA Ⅱ-Ⅲ and left ventricular ejection fraction (LVEF)<40% were enrolled. Based on routine drug therapy, the patients were randomly assigned into 2 groups: Control group, n=50 and Rehabilitation group, n=46, the patients performed treadmill exercise, the training intensity was tailored by (50-80) % of peak oxygen uptake (peak VO2) of baseline cardiopulmonary exercise test (CPET) at (25-40) min/session, 3 sessions/week for 12 weeks. The peak VO2, VE/VCO2 slop, anaerobic threshold (VO2 AT), maximum workload and maximum exercise time were measured by CPET; left atrial diameter (LAD), left ventricular end diastolic diameter (LVEDD), cardiac index (CI) and LVEF were examined by echocardiography; 6 min walking distance (6MWD) and plasma NT-proBNP level were recorded; QoL was assessed by Minnesota living with heart failure questionnaire (MLWHFQ). The above indexes were compared within Rehabilitation group and between 2 groups. Results: In Rehabilitation group, compared to baseline condition, the following indexes were increased by 12 weeks training: peak VO2 (19.8±2.7) ml/min?kg vs (17.4±2.1) ml/min?kg, VO2 AT (11.6±2.5) ml/min?kg vs (9.5±1.8) ml/min?kg, maximum workload (120±20) w vs (102±21) w, maximum exercise time (8.2±1.7) min vs (6.4±1.5) min, CI (2.2±0.5) L/(min?m2) vs (1.9±0.4) L/(min?m2), LVEF (42±5) % vs (35±4) % and 6MWD (406±58) m vs (345±79) m, all P<0.05; while the following parameters were decreased: VE/VCO2 slop (31.7±4.6) vs (34.2±5.8), LAD (38.6±5.5) mm vs (41.5±3.6) mm, LVEDD (58.4±6.3) mm vs (62.9±5.4) mm, NT-proBNP (235±69) ng/ml vs (387±57) ng/ml and MLWHFQ (30.8±12.0) vs (42.3±8.5), all P<0.05. The above indexes were different between Control group and Rehabilitation group, all P<0.05. Conclusion: Rehabilitation may safely and effectively improve cardiopulmonary function and quality of life in CHF patients.

Objective To analyze three different classification criteria, the clinical characteristics of antiphospholipid syndrome(APS)in a cohort of Chinese patients. Methods From January 1996 to October 2006, APS patients diagnosed with different classification criteria were retrospectively studied. Results There were totally 120 APS patients fulfilled at least one criterion, One hundred and one patients fulfilled the 1988 Asherson criteria, 96 patients fulfilled the 1999 Sapporo criteria, and 115 patients fulfilled the 2006 Sydney criteria. The ratio of male to female in a cohort of 115 definite APS patients was 1 to 10.5. The mean period of the disease until entry into the study was 82.6 months, the mean age at study entry was(41?12)years. Ninety patients had thrombosis episodes, among which the most common presenting manifestations were deep venous thrombosis, stroke and skin vasculitis. Forty-six of 92 married women in our cohort had fetal morbidity. Catas- trophic APS occurred in 7 patients. The presence of anticardiolipin antibodies(aCL)was detected in 86 pa- tients, anti-beta-2 glycoproteinⅠantibodies in 58 patients and lupus anticoagulant(LA)in 27 patients. Conclusion The most common presenting manifestations are deep venous thrombosis, stroke and cutaneous manifestations. The sensitivity of Sydney classification criteria is improved by adding anti-beta-2 glycopreteinⅠantibody as one of the laboratory criteria. However, primary APS patients who only presented with thrombo- cytupenia and positive laboratory tests could not satisfy this criterion. In addition, the significance of autoanti- bodies to some coagulant factors in APS needs further study.

Objective To observe the possible association of serum adiponectin level with testosterone level in patients with isolated hypogonadotropic hypogonadism(IHH).Methods The diagnosis of IHH was made,based on clinical presentations,laboratory examinations of hormones and GnRH stimulation test.Serum adiponeetin and testosterone levels were measured in 23 IHH patients and 15 normal men.Results After matched for weight,blood pressure and waist-to-hip ratio,serum testosterone level in IHH group was significantly lower than that in control group [(0.23?0.18 vs 4.20?1.90)?g/L,P

BACKGROUNDThe form deprivation (FD) reduces spatial contrasts and induces myopia. Nitric oxide and cyclic guanosine monophosphate (cGMP) are involved in visual signal transmission. This study investigated changes in nitric oxide synthase (NOS) activity and cGMP concentration in ocular tissues in acute and chronic form deprivation myopia.

METHODSGuinea pigs had one eye covered by translucent glass for 7, 14 or 21 days. Untreated litter mates were used as controls. NOS activity and cGMP concentrations in the retinal, choroidal and scleral tissues of FD eyes and control eyes were analyzed by radioimmunoassay after various durations of FD. The expression of NOS subtypes was identified by immunohistochemistry.

RESULTSMyopia was successfully induced in FD eyes after 14 days. Compared with control groups, the retinal NOS activity and cGMP concentrations in the FD eyes significantly increased after 14 and 21 days while the retinal NOS activity in the FD eyes was transiently suppressed by 7 days of FD. The NOS activity and cGMP concentrations of choroid and sclera in the FD eyes were higher than in the control groups at 21 days. The three isoenzymes of nitric oxide synthase were detected in the ocular tissues of guinea pigs.

CONCLUSIONSThe NOS activity and cGMP concentrations were upregulated after chronic FD and the retinal NOS activity was transiently suppressed at acute FD. The function of elevated NOS activity may be mediated by cGMP.

Animals ; Cyclic GMP ; analysis ; Guinea Pigs ; Immunohistochemistry ; Myopia ; metabolism ; Nitric Oxide ; physiology ; Nitric Oxide Synthase ; metabolism ; Refractive Errors ; Retina ; metabolism