Airway Remodeling ; drug effects ; Cholinergic Antagonists ; therapeutic use ; Humans ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; immunology ; metabolism ; Scopolamine Derivatives ; therapeutic use ; Tiotropium Bromide

Rat calcineurin (CaN) A alpha isoform (Ppp3ca) cDNA recombinant adenovirus vector was constructed in order to explore the effect of CaN on the myocardium apoptosis induced by ischemia-reperfusion injury. Total RNA was isolated from the heart of the adult Wistar rat, and Ppp3ca CDS segment of approximate 1.59 kb size was amplified by reverse transcriptional PCR method. Ppp3ca cDNA segment was cloned into pMD18-T Simple vector for sequencing, and the right clone was named T-Ppp3ca. Ppp3ca cDNA segment obtained from T-Ppp3ca was ligated with pShuttle2-IRES-EGFP to construct a recombinant plasmid pShuttle2-Ppp3ca-IRES-EGFP. Ppp3ca-IRES-EGFP expression cassette containing CMV, Ppp3ca-IRES-EGFP and SV40 polyA DNA fragment (3.97 kb) obtained from pShuttle2-Ppp3ca-IRES-EGFP was connected with pAdeno-X backbone sequence to construct a recombinant plasmid pAdeno-Ppp3ca. After being identified by PCR and enzyme digestion, recombinant plasmid pAdeno-Ppp3ca was packaged in HEK293 cells. Supernatant of adenovirus from HEK293 cells was collected after a visible cytopathic effect (CPE) appeared. The DNA of the recombinant adenovirus was extracted with the standard method. The presence of the recombinant adenovirus was verified by PCR. The results showed that sequencing results verified that the PCR product of Ppp3ca gene was identical to GenBank. Agarose electrophoresis showed the bands of recombined plasmid pAdeno-Ppp3ca and the recombinant adenovirus identified by enzyme digestion and PCR were in the right range corresponding with expectation. It was concluded that the recombinant adenovirus carrying rat calcineurin A alpha (Ppp3ca) cDNA as well as a report gene-enhancer green fluorescent protein gene was successfully constructed in this experiment.
Adenoviridae/*genetics ; Calcineurin/*biosynthesis ; Calcineurin/genetics ; Cloning, Molecular ; DNA, Complementary/genetics ; Genetic Vectors/genetics ; Green Fluorescent Proteins/biosynthesis ; Green Fluorescent Proteins/genetics ; Myocardial Reperfusion Injury/*genetics ; Myocardium/chemistry ; Rats, Wistar ; Recombination, Genetic/genetics

Objective To construct rat calcineurin catalytic subunit ?(CnA)/enhanced green fluorescent protein EGFP gene coexpression plasmid for exploring the effect of calcineurin on the myocardium apoptosis induced by ischemia-reperfusion.Methods Total RNA was isolated from the heart of the adult Wistar rat,and CnA CDS segment of approximate 1 590 bp size was amplified by reverse transcription PCR method.CnA cDNA segment was cloned into pMD18-T Simple vector for sequencing,and the right clone was named T-CnA.CnA cDNA segment excised from plasmid T-CnA was ligated with pShuttle2 which had inserted IRES-EGFP segment into before.The DNA of the recombinant plasmid was extracted and was identified by double digesting with Nhe Ⅰ and Not Ⅰ.The right clone was named pShuttle2-CnA-IRES-EGFP.Results Sequencing result verified that the PCR product of CnA gene was identical to GenBank(NM_017041).1% agarose electrophoresis showed the bands of recombinant plasmid pShuttle2-CnA-IRES-EGFP digested by Nhe Ⅰ and Not Ⅰ were in the right range corresponding with expectation(1 590 bp and 5 240 bp).Conclusion Recombinant eukaryotic expression plasmid carrying rat CnA cDNA as well as a report gene-EGFP gene is successfully constructed in this experiment.

0.05), but hospitalization duration was shorter in the invasive group than in the conservative group (10.3?5.6 days vs 14.6?10.7 days, P

Objective To study the production of ?-lactamase in multidrug-resistant Pseudomonas aeruginosa and to guide the proper use of antibiotics in clinical practice. Methods The modified three-dimensional extract test was employed to detect ?-lactamase in 30 multidrug-resistant Pseudomonas aeruginosa strains screened from antimicrobial susceptibility test in our hospital,and isoelectric focusing electrophoresis was performed on the enzyme-producing strains. Results No metalloenzyme was detected in all the 30 strains.Twenty-six strains produced ?-lactamase,among which 25 continuously yielded large amount of AmpC enzyme and the other one both AmpC enzyme and extended-spectrum ?-lactamases(ESBLs).86.7% of multidrug-resistant Pseudomonas aeruginosa produced enzyme. ConclusionThe majority of the multidrug-resistant Pseudomonas aeruginosa in our hospital yielded large amount of AmpC enzyme in a continuous way.The modified three-dimensional extract test can eliminate some interference such as the decrease of outer membrane permeability and overexpression of efflux pump,facilitating the effective and accurate detection of ESBLs in Pseudomonas aeruginosa.

OBJECTIVE: Analysis of multislice CT (MSCT) on the misdiagnosis reasons of children bronchial foreign body, avoid missed diagnosis, to achieve reasonable application. METHOD: Fourteen cases of misdiagnosed cases of data were retrospectively analyzed in our department of suspicious in children with bronchial foreign body, and discuss the misdiagnosis reasons. RESULT: Fourteen cases of misdiagnosis of children with 9 cases by MSCT examination showed no obvious foreign matter. Through bronchoscopy intraoperative found foreign body, 5 cases by multislice CT (MSCT) to found foreign body, 4 cases of phlegm scabs, 1 case is inflammatory granulation, all recovered after treatment. Reasons of misdiagnosis were threshold selection error, scanning level from the inception glottis, imaging error, etc. CONCLUSION MSCT is a very valuable diagnostic on airway foreign body check method, but there are certain limitations, we should improve the understanding of misdiagnosis and reduce the occurrence of this phenomenon.
Bronchi ; Child, Preschool ; Diagnostic Errors ; Female ; Foreign Bodies ; diagnostic imaging ; Humans ; Infant ; Male ; Retrospective Studies ; Tomography, Spiral Computed ; methods

Objective To discuss the technical specifications of current high-frequency electrotome detection,to avoid the hidden danger of high-frequency electrotome power detectors,and to measure the leakage current of different kinds of highfrequency electrotome accurately.Methods The power and leakage current of the high frequency electrotome were measured by FLUCK QA-ES Ⅱ high frequency electrotome analyzer.The safety of the two methods was compared before and after the improvement of the power measurement.Four parameters of leakage current were repeatedly measured with the ways of high frequency earthing and high frequency isolation respectively.The maximum measurement of leakage current was recorded.Results The improved connection method was safe in the power measurement.For the high-frequency electrotome in the model of high frequency earthing,the values of leakage current were restrained within the range of error with two ways of monopolar loading operation electrode and neutral electrode.For the high-frequency electrotome in the model of high frequency isolation,the values of leakage current were limited within the range of error withtwo ways of monopolar empty operation electrode and neutral electrode.Conclusion The improved high-frequency electrotome power detection method is safe for detectors.The data obtained from the leakage current detection method using the national standard correction method reflect the actual state of the high-frequency electrotome,when the electrotome with earth as the reference is used to detect the leakage current with loading or the insulated electrotome is applied to measuring the leakage current with no loading.

Objective:To investigate the effect of short-term intensive treatment with insulin on metabolic control,?-cell function and insulin resistance in newly diagnosed type 2 diabetic patients. Methods:Thirty-eight newly diagnosed type 2 diabetic patients were randomly divided into two groups and treated with continuous subcutaneous insulin infusion(CSII) or multiple daily insulin injection(MDI) for 1 month.Fasting blood glucose(FBG) and lipid profiles were measured,and oral glucose tolerance test(OGTT)was performed before and after the treatment.The second phase insulin and C peptide secretions were measured by calculating area under curve(AUC) in OGTT.HOMA model was used to evaluate ?-cell function((HOMA-?)) and insulin resistance. Results:Excellent control of FBG was achieved after 1 month intensive treatment.(HOMA-?) was improved significantly. Conclusion:The excellent metabolic control and ?-cell function improvement can be achieved by short-term intensive insulin therapies like CSII or MDI.

Objective To evaluate and monitor the efifcacy of GnRH analogs (GnRHa) therapy. Methods Thirty girls with central precocious puberty diagnosied by LHRH stimulation test were treated with GnRHa for 6-24 months. The LHRH stimula-tion test were performed again at 3 months after initiation of therapy and then every 6 months during treatment. The relationship of peark LH and clinical suppressing pubertal (including Turner stage, bone age, grwoth speed) were compared. The monitor effect of peak LH to efifcacy of GnRHa were eveluated. Results Ninety LHRH stimulation tests were performed, only 7 cases were found to have clinical pubertal development. After 6 months treatment, the base LH level of thirty girls (0.48±0.20) IU/L was signiifcantly lower than that before the treatment (0.75±0.35 IU/L) (P=0.000). The correlation coefifcient between base LH and peak LH was 0.62. The best correlation between clinical suppressing pubertal and LHRH stimulation test was achieved when peak LH was less than 2 IU/L (85.7%sensitivity, 100%speciifcity). Conclusions Base LH value can be used in preliminary as-sessment of the efifcacy of GnRHa therapy for girls with central precocious puberty. The peak LH less than 2 IU/L can be as the indicator of treatment efifcacy.