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.

OBJECTIVETo observe the effects of acupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) on distribution taxis of paclitaxel in mice with lung cancer to discuss targeted relationship between acupoints and corresponding viscera.

METHODSAccording to randomized digital table, 315 SPF-grade BALB/C female mice were divided into 7 groups: blank group (group A), model group (group B), medication group (group C), acupuncture at non-acupoint group (group D), acupuncture at Feishu group (group E), acupuncture at Lingtai group (group F) and acupuncture at Feishu and Lingtai group (group G), 45 mice in each one. Except the blank group, the remaining groups were treated with N-nitroso-tris-chloroethyl urea (NTCU) to establish the model of squamous-cell carcinoma. After model establishment, group A, group B and group C were not treated with acupuncture; group A and group B were treated with intraperitoneal injection of 0.9% sodium chlorvde solution by 6 mL/kg while group C was treated with intraperitoneal injection of paclitaxel by 6 mL/kg. The group D, group E, group F and group G were treated with acupuncture at non-acupoint, "Feishu" (BL 13), "Lingtai" (GV 10) and "Feishu" (BL 13) plus "Lingtai" (GV 10), respectively, then were intraperitoneally injected with paclitaxel by 6 mL/kg. The treatment was all given once a day for continuous 10 days. 15 min, 30 min, 1 h, 2 h, 8 h, 12 h and 24 h after the treatments, 6 mice in each group were randomly selected and sacrificed to collect samples of lung, liver, spleen, kidney and heart, etc. High performance liquid chromatography was applied to measure the concentration of paclitaxel in each organ (lung, liver, spleen, kidney and heart) at different time points.

RESULTS(1) The content of paclitaxel in lung, kidney and heart reached the peak at 2 h, then decreased significantly in group C, group D, group E, group F and group G; the content of paclitaxel in spleen showed downtrend at each time point. The content of paclitaxel in liver reached the peak at 2 h in group C and group D; the content of paclitaxel reached the peak at 8 h in group E, group F and group G. (2) The content of paclitaxel in lung in group E and group G was higher than that in group C and group D at each time point (all P < 0.01); the content of paclitaxel in lung in group F was higher than that in group C (P < 0.01) and group D (P < 0.01) only at time point of 2 h. The content of paclitaxel in lung in group G was higher than that in group F at each time point (all P < 0.05). There was no statistical difference between group G and group E (all P > 0.05).

CONCLUSIONAcupuncture at "Feishu" (BL 13) and "Lingtai" (GV 10) could influ- ence the metabolism of paclitaxel in lung-cancer mice, leading to distribution change in each organ. As a result, it could cause targeting effects, which is more significant at "Feishu" (BL 13) and "Lingtai" (GV 10).

Acupuncture Points ; Acupuncture Therapy ; Animals ; Antineoplastic Agents, Phytogenic ; pharmacokinetics ; Drugs, Chinese Herbal ; pharmacokinetics ; Female ; Humans ; Lung Neoplasms ; drug therapy ; therapy ; Mice ; Mice, Inbred BALB C ; Paclitaxel ; pharmacokinetics ; Taxus ; chemistry

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.

Objective To investigate the bond strengths of customized titanium bracket manufactured by selective laser melting.Methods Eighty 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.Results The 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.Conclusions The shear bond strength and tensile bond strength of both kinds of brackets were enough for clinic application.

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.

Objective To study on the effects of freeze -dry powder of Sini San on sleep in rats.Methods Rats were randomly divided into two groups (freeze -dry powder of Sini San and blank groups )of 15 each day to ensure the alternating light and dark 12 h.The postoperative recovery after two days, rats were continuously administered orally by freeze -dry powder of Sini San 4.95 g? kg-1 for 7 days.Blank group was given dis-tilled water by intragastric way.The other disposal methods were the same with administration group.The efficacy of Sini San on sleeping phase of normal and insomnious rats are carried out by electroencephalo -gram (EEG).Results The total sleeping time (TST) of rats were sig-nificantly prolonged induced by pentobarbital sodium , especially slow -wave sleep 2 (SWS2) and rapid -eye -movement sleep (REMS) were prolonged after the rats were orally administrated of Sini San freeze -dry powder for seven days.The result indicated that Sini San can significant effect on improving sleep.Conclusion There was the function of Sini San in the treatment for insomnia.

Objective To study the pharmacodynamic material basis of licorice andimprove the role of sleep.Methods Thirty Wistar rats were divided into 3 groups, which were normal control group, licorice powder group, glycyrrhetinic acid group.The freeze-dried powder 20 g, water dissolved into 0.5 kg· L-1 solution, administered orally, the normal con-trol group were given equal volume of distilled water.One times a day, continuous 7 d.Blood transitional component were analzed by HPLC method.Results Glycyrrhetinic acid could make the peak area of endog-enous substances in cerebrospinal fluid ( CSF ) higher than the peak area of the blank, the peak area ratio of glycyrrhizin could make the endoge-nous substances in cerebrospinal fluid in the area is about 8.6 times peak area of the blank CSF.That the main chemical components of licorice glycyrrhetinic acid can significantly promote the secretion of endogenous substances in the cerebrospinal fluid, so as to improve the role of sleep.Conclusion By analysis and comparison, which indicates that the active constituent( glycyrrhetinic acid) in licorice are confirmed and could im-prove the sedative function.

Objective To study on sleep improving mechanism of Si Ni San Freeze -dry Powder.Methods Wistar rats were divided into four groups, each group was 10.Which were normal group, Si Ni San powder group, Radix Paeoniae Alba group.Paeoniflorin group.The freeze-dried powder 20 g, water dissolved into 0.5 kg? L-1 solution, administered orally, the normal group were given equal volume of distilled water,1 times a day, continuous 7 d.Blood transitional component were analzed by HPLC method.Results By the analysis of the compound in cerebrospinal fluid, the endogenous substance in cerebrospinal fluid was increased obvi-ously.Increasing peak area of the endogenous substance in cerebrospinal fluid were about 12.5 times the blank′s in cerebrospinal fluid.Conclusion Paeoniflorin in Si Ni San Freeze-dry Powder improves sleep by promo-ting the secretion of 5-serotonin in cerebrospinal fluid.

OBJECTIVETo investigate the mechanical changes of the degenerated lumbar disc with finite element analysis.

METHODSA three dimensional finite element model of a human lumbar spine at the L3-L4 disc was established by the software MIMICS and ABAQUS based on computer tomography images. Degeneration was modeled by changes in geometry and material properties. The model was loaded with 0.3 MPa in axial plane. The von Mises stress on the annulus fiber, nucleus pulposus, endplate and facet joints in healthy and degenerated discs was compared.

RESULTCompared with healthy discs, the von Mises stress of disc distributed in the side of annulus fiber, the stress of nucleus pulposus decreased remarkably, the stress of endplate distributed in the posterior part, and the stress of facet joints increased for the degenerated disc.

CONCLUSIONThe finite element models can provide a method of understanding the relationship between biomechanical performance of the disc due to disc degeneration.

Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Intervertebral Disc ; Intervertebral Disc Degeneration ; diagnosis ; Lumbar Vertebrae ; Stress, Mechanical ; Tomography, X-Ray Computed

OBJECTIVETo observe the function of gamma delta T lymphocytes and the polymorphism of T cell receptor V delta chain in the lungs of asthmatic patients and explore the role of gamma delta T cells in airway inflammation.

METHODSBronchoalveolar lavage fluid BALF was obtained from 7 asthmatic patients and 7 healthy control individuals. The percentage of gamma delta T cell in BALF was measured by flow cytometry. The gamma delta T cell in BALF was purified by magnetic labeled beads. Proliferous activity was examined by MTT assay. Cytokines secreted by gamma delta T cells in medium was assessed by enzyme-linked immunosorbent assay. Polymorphism of T cell receptor V delta chain was detected by RT-PCR and gene scan analysis.

RESULTSThe proportion of gamma delta T cell in the BALF of asthmatic patients [(6.39+/-0.71)%] was significantly higher than that in control subjects [(2.62+/-0.37)%] (P<0.01). The proportion of macrophage in the BALF of asthmatic patients [(81+/-4)] was significantly lower than that in control subjects [(86+/-2)] (P<0.05). The proliferation rate of asthmatic patients [(284.2+/-43.6)%] was significantly higher than that of control subjects [(217.5+/-59.5)%] (P<0.05). Interleukin-4 secreted by gamma delta T cells of asthmatic patients [(18.9+/-3.1) pg/ml)] significantly increased when compared with the control subjects [(14.1+/-3.0) pg/ml] (P<0.05). The polymorphism of T cell receptor V delta chain was not significantly different between these two groups.

CONCLUSIONSThe increase of gamma delta T cells in the lung of asthmatic patients further exacerbates Th1/Th2 disturbance and airway inflammation. Antigen recognition by gamma delta T cells is non-specific.

Adult ; Asthma ; genetics ; immunology ; Bronchoalveolar Lavage Fluid ; cytology ; Case-Control Studies ; Cell Proliferation ; Cytokines ; metabolism ; Female ; Genes, T-Cell Receptor delta ; genetics ; Genes, T-Cell Receptor gamma ; genetics ; Humans ; Immunoglobulin Variable Region ; genetics ; Lung ; immunology ; Male ; Middle Aged ; Polymorphism, Genetic ; T-Lymphocyte Subsets ; immunology ; metabolism ; Th1-Th2 Balance