OBJECTIVE To perform a surveillance study in order to investigate the change in antimicrobial resistance of Klebsiella pneumoniae,especially extended-spectrum ?-lactamases(ESBLs)-producing strains isolated.METHODS A total of 1986 strains of K.pneumoniae isolated during the last six years were analyzed.RESULTS Among 1986 K.pneumoniae strains,908(45.7%) were ESBLs-producing strains.The sensitive rate of ESBLs-producing K.pneumoniae to imipenem was 100%,and to meropenem was 99%.There were 636 strains isolated from ICU,accounted for 32%;and 480 strains isolated from Department of Respiratory Medicine,accounted for 24.2%.There were 1630 strains isolated from sputum,accounted for 82.1%.CONCLUSIONS K.pneumoniae is the major pathogen.Detecting ESBLs-producing strains rate and their susceptibility to antibiotics is very important in guiding the clinical administration of drugs.

Objective To construct a phage antibody Fab library of human antinuclear antibody. Methods Total RNA was extracted from peripheral blood lymphocytes of 4 patients suffering from autoallergic disease, with antinuclear IgG antibody titers in the serum higher than 1∶10 000 as estimated by indirect immunofluorescence technique. Taking oligo-dT as the primers, human immunoglobulin light chains ?/? genes as well as heavy chains Fd genes were reversely transcribed to cDNA by PCR. The ?/? light chains were initially cloned into pComb3Hss vector to construct a human recombinant light chain library, and the heavy chain genes were subsequently inserted into the corresponding sites of ?/?-pComb3Hss plasmid to generate a recombinant ?/?-Fd-pComb3Hss plasmid. The plasmid was then transformed into E. coli XL1-Blue, which was subsequently infected by the helper phage M13KO7. A random recombinant antibody library was expressed on the surface of filamentous phage. Results Human immunoglobulin ?/? light chain and heavy chain Fd genes (660bp) were amplified successfully. The light chain library with the capacity size of 2?104, and the human recombinant Fab antibody library with the capacity size of 4?104 were also successfully obtained. Finally, the phage antibody library of the Fab fragment of human antinuclear antibody, containing 2?109 CFU/ml phage, was constructed. Conclusion A phage antibody library of Fab fragment of human antinuclear antibody is constructed successfully. This research lays a foundation for the preparation for phage library of Fab fragment of human antinuclear antibody, and also paves the way for the advanced assessment for its effect on the immuno-targeting therapy of tumors.

Objective To establish a system in which mouse embryonic stem cells (mESCs) differentiate into insulin-secreting cells in vitro. Methods mESCs were cultured to form embryoid bodies(EBs). EBs were cultured in serum-free medium to obtain nestin-positive cells. The selected nestin-positive cells were expanded,then added nicotinamide into the medium to induce the differentiation of nestin-positive cells. Immunochemistry staining and flow cytometry analysis were made on 15th day after induction.Results Flow cytometry analysis revealed that the percentage of nestin-positive cells were different from EBs. The percentage of nestin-positive cells from EBs were higher than those of other diameters. Nestin-positive cells induced by nicotinamide formed islet-like cell clusters. Percentage of insulin-positive cell induced by 10 mmol/L nicotinamide was higher than those induced by 0 mmol/L or 5 mmol/L nicotinamide(P

Objective To summarize the experience of endoscopic thyroidectomy for hyperthyroidism.Methods Endoscopic total or subtotal thyroidectomy was performed through anterior chest wall approach in 7 patients with primary or secondary hyperthyroidism.Results The operation was successfully performed in all the 7 patients.The operation time was 130~260 min(mean,168 min),and the intraoperative blood loss was 10~200 ml(mean,70 ml).No recurrent laryngeal nerve or superior laryngeal nerve injuries,or postoperative hemorrhage,or conversions to open surgery were encountered.The postoperative recovery was uneventful.Short-term follow-up observations demonstrated satisfactory cosmetic results and no recurrence.Hypothyroidism occurred in 2 patients and thyroid functions restored to normal levels in 1 of them 2 months after operation.Conclusions Endoscopic thyroidectomy is a safe and effective procedure for hyperthyroidism.Apart from conventional pre-operative preparation,CT examination is also necessary for identifying the measurements of thyroid glands and determining the proportion and location of residual glands.

OBJECTIVE:To observe the short-term efficacy and safety of nimesulide combined with ibuprofen in the treatment of pain. METHODS:84 patients with pain were randomly divided into observation group and control group. Control group was oral-ly given 0.3 g Ibuprofen sustained-release capsule,twice a day. Observation group was additionally given 100 mg Nimesulide cap-sule,twice a day. The treatment course for both groups was 7 d. Clinical efficacy,visual analogue (VAS) score,quality of life (QOL) score,coagulation indexes (prothrombin time,thrombin time,fibrinogen,activated partial thromboplastin time) before and after treatment,and incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observa-tion group was significantly higher than control group,the differences were statistically significant(P<0.05). Before treatment, there were no significant differences in the VAS score and QOL score between 2 groups(P>0.05). After treatment,VAS scores in 2 groups were significantly lower than before,and observation group was lower than control group,QOL scores were significantly higher than before,and observation group was higher than control group,the differences were statistically significant(P<0.05). And there were no significant differences in the coagulation indexes and incidence of adverse reactions between 2 groups(P>0.05). CONCLUSIONS:The short-term efficacy of ibuprofen combined with nimesulide is superior to ibuprofen alone in the treatment of pain,with similar short-term safety.

Objective To observe the effect of sesamol on the hematopoietic system in mice exposed to 4 Gy irradiation. Method Twenty C 57 BL/6 mice were randomly divided into control group, sesamol group, irradiated group and irradiated+sesamol group (n=5). Mice of control and sesamol group received sham irradiation, and the rest exposed to 4 Gy total body irradiation, dose rate 1.01 Gy/min. Mice in sesamol group and irradiated+sesamol group received a dose of 10 mg/kg sesamol administered by gavage every day for 7 days after irradiation exposure. Mice of other two groups were treated with vehicle solution. After 4 Gy irradiation 7 day, the peripheral bloods were collected. The levels of colony forming units-granulocyte-macrophage (CFU-GM) were detected. Results Compared to irradiation group, the level of WBC、cell count of BMNCs and CFU-GM significantly decreased in the irradiated mice, decreased in the irradiated mice (P<0.05). Compared to irradiation group, cell count of BMNCs and CFU-GM in the irradiated+sesamol group increased significantly (P<0.05). Conclusion Sesamol has a certain impact on the radiation-induced changes in hematopoietic system. The mechanism need to be further explored.

Objective To observe the different radiosensitivity induced by different doses of 137Csγ-ray irradiation between hematopoietic stem and progenitor cells. Methods Seventy-two C57BL/6 mice were randomly divided into control group and irradiated groups (2, 4 and 6 Csγ-ray irradiation, n=18 for each group). Mice of control group received sham irradiation, and the rest accepted 2, 4 and 6 Gy137Csγtotal body irradiation, respectively. After 14-day, 35-day and 56-day irradiation, the peripheral blood samples were collected by balls enucleation. The number of bone marrow nuclear cells, hematopoietic stem and progenitor cells were counted. Results The peripheral blood of irradiated mice showed significant changes in the number of white blood cells (WBC), red blood cells (RBC), platelets (PLT) and hemoglobin (HGB) in a dose-response relationship. Compared with the control group, the numbers of BMNCs and hematopoietic progenitor cells (HPCs) were significantly lower in irradiated group. At 35 d and 56 d after 6 Gy irradiation the numbers of BMNCs and HPCs were significantly lower than those of control group (P<0.05). There were no significant differences in numbers of BMNCs and HPCs between irradiated groups (2 and 4 Gy) and control group. The number of bone marrow hematopoietic stem cells (HSCs) was significantly lower in irradiated group than that in control group after 14-d and 56-d irradiation (P<0.05). Conclusion 137Csγ-ray irradiation has some damage in mouse hematopoietic system. The damage caused by radiation is persistent to hematopoietic stem cells.

Objective To investigate the protective effect of sesamol on radiation injury mouse bone marrow c-kit+cell,and further explore its possible mechanism.Methods Mouse bone marrow c-kit+cells were collected by immunomagnetic cell sorting method.There were 2 groups in the study:single dosing group and radiation plus drug group(doses of irradiation included 1 Gy and 4 Gy),and 10 -8 ~10 -3 mol/L sesamol were co-cultured with mouse bone marrow c-kit+cell half hour before irradiation exposure,cells were then cultured for 18 hours under the conventional culture conditions (37℃ and 5% CO2 ).The viability of mouse bone marrow c-kit+cells were measured by bioluminescence.The ability of colony-forming units were detected by CFU-GM and apoptotic rate of c-kit+cells were detected by Annexin V/PI antiapoptotic assay. Results Compared with control group,after 1 Gy and 4 Gy irradiated,cell viability of mouse bone marrow c-kit+cells were decreased 59.52% and 79.35%,respectively(P<0.05),the number of colony-forming were decreased 40.38% and 87.69%,respectively(P<0.05 ).Cell viability of c-kit+cells and the number of colonies formed were significantly increased with sesamol concentration between 10 -8 ~10 -6 mol/L,but not improve apoptosis rate.Conclusion Sesamol has protective effect on irradiation-induced injury in mouse bone marrow c-kit+cells,the mechanism of which may be related to the ability of hematopoietic progenitor cells proliferation.

Objective To observe the protective effect of N-acetyl-cysteine (NAC) on the injury of irradiation-in-duced bone marrow mononuclear cells (BMMNCs), and explore the possible mechanism. Methods There were 3 groups in the study:control group, irradiation group (doses of irradiation were 1 Gy and 4 Gy) and irradiation with NAC group (NAC was cocultured with BMMNCs half hour before irradiation). The 2×106/mL BMMNCs and the RPMI-1640 medium or 2×10-5 mol/L NAC were added into the 2 mL EP tubes according to the different requirement of groups. The tubes were then cul-tured in the 37℃CO2 incubator for 30 min and irradiated with 1 Gy and 4 Gy. The viability of BMMNCs was measured by bioluminescence. The level of reactive oxygen species (ROS) was measured by DCFH-DA, and the ability of colony-forming units was detected by CFU-GM. Results After 4 Gy irradiation exposure, the cell viability of BMMNCs was significantly lower in irradiation group (284 296.7±16 541.2) than that of control group (848 586.7±61 404.4). After 1 Gy irradiation expo-sure, the level of ROS was higher in irradiation group (6 750.0±103.5) than that of control group (5 710.7±56.2). The number of colony-forming units per 105 cells after irradiation exposure was (626.7±51.3), which was significantly lower than that of control group (986.7±100.7). Compared to irradiation group, the cell viability of BMMNCs increased to (352 770.0±23 466.1) in irradiation with NAC group. The level of ROS decreased to (5 430.0±61.0), and the number of colony-forming units per 105 cells increased to (773.3 ± 49.3). Conclusion NAC has protective effect on irradiation-induced injury in BMMNCs, which may be related with the decreased level of ROS. NAC can play the role of positive control for the following work.

Objective To observe the injury effect of ionizing radiation on bone marrow derived c-kit+ cells.Methods Via-bility of c-kit+ cells was measured by bioluminescence;the level of c-kit+ cells reactive oxygen species was measured by DCFH-DA, the ability of colony-forming units was reflected by CFU-GM;proliferation and apoptosis of c-kit+ cells were measured by flow cy-tometry;the variation of pathway was detected by arrays of gene chip.Results Compared to control group(0 Gy).It had a decrease of c-kit+ cells′cell viability and the ability of colony-forming units after the cells receipt irradiation with the dose of 1 Gy and 4 Gy;and the level of cell reactive oxygen species,ratio of apoptosis cells increased.After 1 Gy irradiation exposure,the ratio of prolifera-tion(S/G2/M phase)cells increased compared to control group.However,when the c-kit+ cells were receipt 4 Gy irradiation expo-sure,the ratio of proliferation(S/G2/M phase)cells decreased.After 4 Gy irradiation exposure,the up-regulate genes contained Srxn1,Psmb5,Cdkn1a,Smc1b,Bcl2l1,Lrdd and so on;the down-regulate genes contained Mpo,Mtf1,Chek1,Rcc1 Ebag9,Ciapin1 and so on.Conclusion There was injury effect of ionizing radiation on c-kit+ cells,and it could induce variation of many pathways.