Abscess ; therapy ; Adult ; Aged ; Catheterization ; methods ; Drainage ; methods ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed

Objective To investigate the resistant mechanism of imipenem-resistant K. pneumoniae.Methods The minimal inhibitive concentrations (MICs) of the antimicrobial agents were determined by Etest.Isoelectric focusing electrophoresis (IEF),plasmid extraction,conjugation, transformation,PCR amplification,cloning and sequencing were carried out for analyzing the encoding gene of ?-1actamases.Results Three kinds of ?-1actamases were detected with pIs of 7.2,6.7,and 5.4.in a clinical strain of K.pneumoniae.These ?-1actamases were TEM-I (pI,5.4),SHV-12 (pI,8.2) and KPC-2 ( pI,6.7 ) confirmed by sequencing of the PCR products.Only one band of ?-1actamase with pI 6.7 was displayed in the transformant.A 1500 bp segment,which contained the KPC-2 gene confirmed by nucleotide sequence analysis,was cloned from a 60 000 bp plasmid of the transformant.Conclusion The strain of K.pneumoniae resistant to imipenem produces a plasmid-mediated carbapenemase KPC-2 which belongs to Bush group 2f,class A ?-1actamase.

A strain was isolated from internal organ of died porcine about 8 weeks with purulent pneumonia,arthritis,pyogenic arthritis and endocarditis in April 2007.Objectives of the study are to confirm the genus of the strain,pathopoiesis,and drug sensitivity.The mainly study methods:the first,the strain was identified by the phenotype and the characteristics of the biochemistry,sequence 16S rDNA genes of the strain was analyzed by molecular biology technology,finally animal experiment and drug sensitivity testing were done.The results of the phenotype and the characteristics of the biochemistry showed that it is greatly similar to Actinomyces hyovaginalis,16S rRNA sequence analysis exhibited the homology achieved to 99.2% com-pared with group III strains of Actinomyces hyovaginalis,and the phyletic evolution analysis also indicated that it has mostly relationship with group III strains of Actinomyces hyovaginalis.Animal experiment dis-covered it has highly pathogenicity to Mus musculus albus;Drug sensitivity testing showed that it is hyper-sensitive to Erycin,Gentamicin and Amikacin.So,the result of the study confirmed that the strain is Actin-omyces hyovaginalis III with the pathogenicity.

Objective To monitor nosocomial infections of Klebsiella pneumoniae by analyzing the randomly amplified polymorphic DNA (RAPD). Methods RAPD analysis was used to genotype 72 Klebsiella pneumoniae strains isolated from the intensive care unit (ICU) of the neurosurgical department, and the fingerprints were compared to confirm the outbreak of nosocomial infection of Klebsiella pneumoniae. Results 72 strains Klebsiella pneurnoniae were classified into 68 RAPD types, suggesting the absence of an outbreak of nosocomial infection of glebsiella pneurnoniae in the ICU.Conclusian RAPD genotyping can be used in the molecular epidemiological study ofKlebsiella pneurnoniae.

OBJECTIVETo investigate the bond strengths of customized titanium bracket manufactured by selective laser melting.

METHODSEighty human premolars which had been extracted for orthodontic purpose were collected and divided randomly (by random table) into two groups (customized bracket group and 3M bracket group, 40 molars in each group). The 35% phosphoric acid was used for etching and the brackets were bonded with 3M Unitek bonding adhesive. All bonded specimens were placed in saline for 24 hours at room temperature and were tested on DWD3050 electronic testing machine to determine the shear bond strength and tensile bond strength. After debonding, the adhesive remnant indexes (ARI) were recorded.

RESULTSThe shear bond strengths of customized brackets was 6.80 (6.20, 8.32) MPa, which was significantly lower than that of the 3M brackets [10.46 (9.72, 11.48) MPa] (Z = -3.463, P < 0.05). And the tensile bond strengths of customized brackets was (6.93 ± 1.21) MPa, which was significantly higher than that of the 3M brackets [(5.88 ± 1.23) MPa] (t = 2.81, P < 0.05). No significant difference was found in the ARI between two different kinds of the brackets.

CONCLUSIONSThe shear bond strength and tensile bond strength of both kinds of brackets were enough for clinic application.

Acid Etching, Dental ; Adolescent ; Bicuspid ; Child ; Composite Resins ; chemistry ; Dental Bonding ; methods ; Dental Debonding ; Dentin-Bonding Agents ; chemistry ; Humans ; Lasers ; Orthodontic Brackets ; Phosphoric Acids ; chemistry ; Random Allocation ; Shear Strength ; Tensile Strength ; Titanium

OBJECTIVETo study the protective effect of Shenxiong injection on the cerebral ischemia-reperfusion injury of senile rats.

METHODTotally 108 Sprague-Dawley (SD) rats were randomly divided into the sham operation group, the model group, the Ni-modipine group and Shenxiong injection groups (low, middle, and high doses). The rat brain ischemia-reperfusion model was established by the middle cerebral artery occlusion (MCAO) method in rats, in order to observe the effect of Shenxiong injection on neurological score and brain infarct volume of rats with cerebral ischemia-reperfusion injury, and determine the contents of NOS, NO, SOD, MDA and LDH in brain tissues. The contents of TNF-alpha and IL-1beta levels in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA) method.

RESULTShenxiong injection could significantly decrease neurological score, injury degree of brain tissues and brain infarct volume of rats with cerebral ischemia-reperfusion injury, increase the vigor of SOD, decrease the levels of MDA, NO, NOS and LDH, and inhibit IL-1beta and TNF-alpha expressions.

CONCLUSIONShenxiong injection has the obvious protective effect on the brain ischemia-reperfusion injury in rats. Its mechanism may be related to the improvement of neurological function, the reduction of free radical injury, and the inhibition of inflammation factor expression.

Animals ; Brain ; blood supply ; drug effects ; metabolism ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Injections ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; drug therapy ; enzymology ; metabolism ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the effect of FK506 on the cavernous nerve regeneration after injury and to discuss its possible action mechanisms.

METHODSFifty-four male adult Sprague-Dawley rats were randomized into three groups: Group 1 (sham control), Group 2 (unilateral cavernous nerve ablation), and Group 3 (unilateral cavernous nerve ablation with subsequent injection of FK506). Electrostimulation of the cavernous nerve was performed at 1 and 3 months after surgical injury. The intracavernous pressure was continuously detected and the rats were followed by nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) staining to identify NOS in the penile nerve fibers of the penile shaft.

RESULTSAt 1 month, the number of NOS-positive nerve fibers significantly decreased with no statistical difference among the three groups except the sham controls (P > 0.05). At 3 months, electrostimulation revealed greater maximal intracavernous pressure in Group 3 than in Group 2 (P < 0.01). Furthermore, the number of NOS-positive nerve fibers showed a significant increase (P < 0.01), but not in Group 2 (P > 0.05).

CONCLUSIONFK506 injection enhances the regeneration of cavernous nerves after injury and the recovery of erectile function in rats.

Animals ; Electric Stimulation ; Male ; Nerve Fibers ; enzymology ; physiology ; Nerve Regeneration ; drug effects ; Nitric Oxide Synthase ; analysis ; Penile Erection ; drug effects ; Penis ; innervation ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tacrolimus ; pharmacology

BACKGROUNDMajority of the research on cardiac arrest (CA) have focused on post-CA brain injury and myocardial dysfunction, the renal dysfunction and acute kidney injury (AKI) in other critical illnesses after CA have not been well described. This study was designed to assess AKI with renal Doppler and novel AKI biomarkers in a swine model of ventricular fibrillation cardiac arrest (VFCA).

METHODSThirty healthy piglets were divided into VFCA group (n = 22) and Sham group (n = 8) in a blinded manner. Mean arterial pressure, heart rate, and cardiac output were recorded continuously. Cardiac arrest (CA) was induced by programmed electric stimulation in the VFCA group, and then cardiopulmonary resuscitation was performed. Twenty piglets returned of spontaneous circulation (ROSC) and received intensive care. Blood and urine samples were collected for AKI biomarkers testing, and Color Doppler flow imaging was performed at baseline, 6 h, 12 h, and 24 h, respectively after ROSC. At ROSC 24 h, the animals were sacrificed and a semi-quantitative evaluation of pathologic kidney injury was performed.

RESULTSIn the VFCA group, corrected resistive index (cRI) increased from 0.47 ± 0.03 to 0.64 ± 0.06, and pulsatility index (PI) decreased from 0.82 ± 0.03 to 0.68 ± 0.04 after ROSC. Cystatin C (CysC) in both serum and urine samples increased at ROSC 6 h, but neutrophil gelatinase-associated lipocalin (NGAL) in serum increased to 5.34 ± 1.68 ng/ml at ROSC 6 h, and then decreased to 3.16 ± 0.69 ng/ml at ROSC 24 h while CysC increasing constantly. According to the renal histopathology, 18 of 20 animals suffered from kidney injury. The grade of renal injury was highly correlated with RI, cRI, NGAL, and CysC. Linear regression equation was established: Grade of renal injury = 0.002 × serum CysC + 6.489 × PI + 4.544 × cRI - 8.358 (r2 = 0.698, F = 18.506, P < 0.001).

CONCLUSIONSAKI is common in post-CA syndrome. Renal Doppler and novel AKI biomarkers in serum and urine are of significant importance as early predictors of post-CA AKI.

Acute Kidney Injury ; blood ; etiology ; Animals ; Biomarkers ; blood ; Cystatin C ; blood ; Disease Models, Animal ; Female ; Heart Arrest ; blood ; complications ; Lipocalins ; blood ; Male ; Swine ; Ultrasonography, Doppler ; methods ; Ventricular Fibrillation ; blood ; complications

To combine the fibrinolytic with anticoagulant activities for therapy of thrombotic deseases, a fusion protein made of tissue-type plasminogen activator (t-PA) and hirudin was constructed and expressed in chia pastoris. To improve thrombolytic properties of t-PA and reduce bleeding side effect of hirudin, FXa-recognition sequence was introduced between t-PA and hirudin molecules.The anticoagulant activity of hirudin can be target-released through cleavage of FXa at thrombus site. t-PA gene and hirudin gene with FXa-recognition sequence at its 5'-terminal were obtained by RT-PCR and PCR respectively. The fusion protein gene was cloned into plasmid pIC9K and electroporated into the genome of Pichia pastoris GS115. The expression of fusion protein was induced by methanol in shaking flask and secreted into the culture medium. Two forms of the fusion protein, single-chain and double-chain linked by a disulfide bond (due to the cleveage of t-PA at Arg275-Ile276), were obtained. The intact fusion protein retained the fibrinolytic activity but lacked any anticoagulant activity. After cleavage by FXa, the fusion protein liberated intact free hirudin to exert its anticoagulant activity. So, the fusion protein is a bifunctional molecule having good prospect to develop into a new targeted therapeutic agent with reduced bleeding side effect for thrombotic diseases.
Animals ; Cloning, Molecular ; Electroporation ; Hirudins ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Plasminogen Activator ; biosynthesis ; genetics

OBJECTIVETo construct effective RNA-interference plasmids targeting mouse HIF-1alpha gene and testify their effects and specificities in interfering HIF-1alpha expression.

METHODSThree RNA-interference plasmids targeting mouse HIF-1alpha gene, pBS/U6/HIF-1alpha-siRNAI~III, were constructed and identified using double digestion method in the present study. RT-PCR, immunostaining and western blotting were employed to detect the expression alterations of HIF-1alpha in 293T cells following transfections of the three plasmids, respectively. The interference effect of pBS/U6/HIF1alphai-II in SH-SY5Y cell line was further investigated.

RESULTSAll the three RNA-interference plasmids, especially pBS/U6/HIF1alphai-II, showed significant inhibition in HIF-1alpha expression in 293T cell line. pBS/U6/HIF1alphai-II could also inhibit HIF-1alpha expression in SH-SY5Y cell line, in a dose-dependent way.

CONCLUSIONPlasmid pBS/U6/HIF1alphai-II constructed in our study can effectively and specifically inhibit HIF-1alpha expression, and its role in neural tube development and dysfunction will be further investigated. Construct of pBS/U6/HIF1alphai-II plasmid will provide a useful tool to study the role of HIF-1 pathway in embryogenesis, oncogenesis and ischemia development.

Analysis of Variance ; Animals ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Gene Silencing ; physiology ; Green Fluorescent Proteins ; genetics ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Mice ; Plasmids ; genetics ; RNA, Small Interfering ; genetics ; pharmacology ; Transfection ; methods