Objective To explore the effect of nuclear factor-kappa B (NF-κB) decoy oligode-oxynucleotides (ODN) on respiratory function and expressions of IL-1β and IL-13 in serum following se-vere lung contusion in rabbits. Methods A total of 40 New-Zealand rabbits were randomly divided into four groups, ie, severe lung contusion group (Group A, n=12), severe lung contusion with NF-κB scrambled decoy ODN intervention group (Group B, n=12), severe lung contusion with sense NF- B de-coy ODN intervention group (Group C, n=12) and normal control group (Group D, n =4). After the contusion model was set up, the sense and scrambled NF-κB decoy ODN were infused into the rabbits via the jugular veins in different groups, with 20 g per experimental rabbit. After contusion, respiratory fre-quency, tidal volume, airway pressure, respiration flow rate curve and end expiration nitric oxide concen-tration were detected at 1, 2, 3 and 4 hours. The expressions of IL-1β and IL-13 in serum were observed by means of ELISA. Results After sense NF-κB decoy ODN intervention, alveolar ventilation, arteri-al PO_2 and pulmonary compliance were improved, compared with Group A and Group B, with statistical difference (P<0.01). The expression of IL-1β was decreased and that of IL-13 increased after sense NF-κB decoy ODN intervention to the severe lung contusion, compared with Groups A and B, with statis-tical difference (P <0.01). The expression of IL-1β was increased to peak level at 1 hour after contu-sion, which continued to the end of the experiment. While expression of IL-13 was decreased at 1 hour af-ter contusion and reached the minimum level at 4 hours. With intervention with sense decoy ODN, the in-creased expression of IL-1β was down-regulated, but expression of IL-13 remained at high level, with sta-tistical difference compared with Group A and Group B (P < 0.01). Conclusions Intervention with sense NF-κB decoy ODN can significantly protect the respiratory function, reduce the expression of IL-1β and increase expression of IL-13 after severe lung contusion.

AIM: to establish the cell lines whose hREV3 gene expression was blocked by antisense RNA and observe their characteristis of cell growth rate and N - methyl - N' - nitrosoguanidine (MNNG) sensitivi- ty . METHODS: With modified calcium phosphate - DNA coprecipitation method the eukaryocytic expression plasmid expressing antisense fragment of hREV3,pBK - RSV - hREV3- and pMAMneo - amp hREV3 were transfected into human embryo kidney cell line of HEK - 293. After G418 selection, cell lines of 293 - B - hREV3- and 293 - M hREV3- were established. By cell counting method, the cell growth rate and MNNG sensitivity of these cell lines were characterized. RESULTS: No change of cell growth rates of these cell lines was observed whether hREV3 gene expres- sion was blocked by either the persistent or induced expression of the antisense hREV3 RNA. While the sensitivity of these cell lines to MNNG was somewhat elevated, as compared with their parent cell line 293 and the cell lines trans- fected with vector DNAs. CONCLUSION: The gene product of hREV3 was not essential for the cell growth, but it may play a role in the DNA repair functions of the cells after exposure to DNA damaging agents.

Objective To investigate the distribution of acquired carbapenemases in carbapenemresistant strains of Klebsiella pneumoniae, and explore its role in epidemiology of nosocomial infection. Methods From November 2008 to March 2009, twenty clinical isolates of carbapenem-resistant Klebsiella pneumoniae were collected from children hospitalized in Wuhan children's hospital. MICs of antibiotics were tested by DNA of Klebsiella pneumoniae. Modified Hodge test was used to screen strains producing carbapenemases,combined imipenem(IPM)-EDTA , meropenem(MEM)- EDTA and ceftazidime(CAZ) - EDTA double-disk synergy test (DDST) were used to detect metallo-β-lactamase-producing. PCR amplification of the carbapenemase and integrase genes, and sequencing were performed. Plasmid conjugation transfer experiments and Southern hybridization were applied to study the mode of drug resistance transmission. Results Four types of Klebsiella pneumoniae were detected by PFGE, type A consisted of 5 strains, including 3 strains of type Al and 2 strains of type A2), type B (2 strains), type C (12strains) and type D (1 strain). Type A and C were the main drug resistant clones. Eight strains of Klebsiella pneumoniae carried both KPC-2 and IMP-4 genes, 10 strains carried IMP-4 gene, 2 strains carried KPC-2 gene. None of NDM-1 ,GIM, SPM, SIM, OXA-23, and VIM carbapenemase genes was detected in 20 isolates. All of 20 isolates carried lntl which were found to be located on bacterial chromosome by Southern blot. Conclusions KPC-2and IMP-4 genes are the major carbapenemase genes in Klebsiella pneumoniae isolated in Wuhan.Transmission of drug resistance is mainly through vertical transmission of type C resistant clone and horizontal transmission of Intl on bacteria chromosome.

Objective To evaluate the effects of external substance P (SP) on scalding wound healing and neovascularization.Methods Eighty-four Wistar rats were induced into diabetic models by intraperitoneal injection of streptozotocin (STZ), and deep partial thickness scalding wound on the back with diameter of 2 cm was prepared. Rats were randomly divided into experiment group (n=42, local injection of SP) and control group (n=42, local injection of PBS). The process of wound healing was observed, and the percentages of wound closure were calculated on D0, D1, D3, D7, D10, D14, D21 post scald. The expression of CD31, surface marker of neovascular endothelial cells, was detected within the wound sites by immunohistochemical staining, and the microvessel density was calculated. Results The percentage of wound closure in experiment group was significantly higher than that in control group on D7 post scald [(42.69±3.26) % vs (30.24±1.17)%, P<0.01]. Immunohistochemical detection revealed that the expression of CD31 and the microvessel density in experiment group were significantly higher than those in control group from D7 post scald (P<0.01). ConclusionExternal SP may promote scalding wound healing in diabetic rats, the mechanism of which may be associated with upregulation of expression of CD31 and acceleration of neovascularization.

Objective To investigate the molecular epidemiology of CRAB isolated from children in wuhan. Methods Forty non-repetitive strains of CRAB were collected from hospitalized children of emergency department, neonatal medicine, cardiothoracic surgery, bone surgery, respiratory medicine and renal medicine in Wuhan children's hospital during December 2008 and May 2009. MIC values were PFGE; KPC, IMP, GIM, SPM, SIM, OXA-23, VIM genes and integrase gene were amplified by PCR and then sequenced to confirm the genotypes.; Plasmid conjugation experiment was used to study the transfer method of bacterial resistance and southern blot hybridization was used to target the resistance genes. Results Susceptible rates of 40 strains to gentamicin, tobramycin, amikacin, ciprofloxacin, levofloxacin, trimoxazole were 20%, 5%, 93%, 93%, 95%, and 23% respectively. Eleven types of clone were detected by PFGE,including 29 strains of type A clone, 2 strains of type B clone, and 1 strain for each type of C to K clone. Eleven isolates produced both IMP-4 and OXA-23 carbapemase. Twenty-six isolates only possessed OXA-23 carbapemase. Thirty-six strains carried class Ⅰ integron. The results of southern blot hybridization showed that Intl, IMP-4 and OXA-23 type were located on chromosome. Conclusions Type A clone of CRAB is the most common. OXA-23 and IMP-4 type are the major acquired carbapemases, especially the OXA-23 is the most common type. The horizontal transmission of OXA-23 and IMP-4 gene mediated by Int1 and the spread of type A resistant clone is the major way of the spread of carbapenem-resistant Acinetobacter baumannii in the region.

Objective:To compare the sedative effect and safety of different doses of dexmedetomidine in the patients undergoing total abdominal hyserectomy. Methods:A total of 120 patients undergoing total abdominal hysterectomy were randomly divided into four groups, different doses of dexmedetomidine groups(D1 group, D2 group and D3 group)and midazolam group(M group) with 30 pa-tients in each. Dexmedetomidine groups received intravenous pump infusion of dexmedetomidine (0. 5 μg·kg-1 ) 10 minutes before the operation, and then the different dexmedetomidine groups were received continuous infusion of dexmedetomine of different doses:D1 group of 0. 4μg·(kg·h) -1, D2 group with 0. 6μg·(kg·h) -1 and D3 group with 0. 8μg·(kg·h) -1;M group received in-travenous pump infusion of midazolam (0. 06 mg·kg-1) 10 minutes before the operation, and then with 0. 04 mg·(kg·h) -1 con-tinuous infusion. The mean arterial pressure( MAP) , heart rate( HR) , respiratory rate( RR) , oxyhemoglobin saturation( SpO2 ) were recorded at the following time points:the moment of entering the operating room(T0), the block effect of epidural anesthesia was satis-fied (T1),10 min(T2),20 min(T3)and 40 min(T4)after the drug infusion, and the end of the operation(T5), and the duration of the medicine use and the whole operation were recorded as well. The sedation degrees were evaluated with Ramsay scale, and the am-nesic scores, adverse drug reactions and patient satisfaction were recorded after the operation. Results:Compared with that of the other groups, HR of D3 group was obviously lower after T3 (P<0. 05);and after T2, HR was significantly lower than that at T0 (P<0. 05 or P<0. 01). Compared with the other groups, RR of M group was obviously lower at T3 and T4 (P<0. 05). Compared with those at T0 , the sedative effects of all the groups were much remarkable(P<0. 05 or P<0. 01);and the Ramsay score of D3 group at T3 and T4 was higher than that in D1 group or M group(P<0. 05). There were no obvious adverse reactions in the four groups. Conclusion:The use of dexmedetomidine in the patients undergoing total abdominal hyserectomy might not lead to the risk of respiratory depression as the use of midazolam, while the dose of dexmedetomidine should be less than 0. 8 μg·(kg·h) -1.

Objective To explore the effect of ILK on myocardium contraction following hemorrhagic shock .Methods With iso-lated cardiac papillary muscle and isolated heart ,the contraction of papillary muscle and hemodynamic parameters (left intraventricu-lar systolic pressure(LVSP) ,the maximal change rate of left intraventricular pressure were measured .Results Compared with nor-mal control hearts ,ILK activity ,contractile response of cardiac papillary muscle and hemodynamic parameter were decreased signifi-cantly gradually in shock heart at the 1 ,2 h(P<0 .05) ,and the change of ILK activity was positive correlative with contractile re-sponse and hemodynamic parametert .With ILK agonist [phosphatidylinositol(3 ,4 ,5)trisphosphate ,PLTP] the dysfunction of con-tractile response and hemodynamic parameter could be improved significantly (P<0 .05) ,while these improvement could be abol-ished by ILK specific inhibitor(P<0 .05) .Conclusion ILK play important role in the regulation of cardiac contractility following hemorrhagic shock .

Animals ; Female ; Pregnancy ; Pyrethrins ; toxicity ; Rats ; Rats, Wistar ; Teratogens ; toxicity

Objective To explore the effect of autophagy regulator on the injury of rat hippocampal neurons induced by oxygen-glucose deprivation (OGD).Methods Rat hippocampal neurons were cultivated in primary and subjected to OGD to simulate neuronal hypoxic ischemia injury for 2 hours or 6 hours followed by reperfusion for 12 hours with or without 3-methyladenine (3-MA, 20μmol/L) or rapamycin (0.2μmol/L). The morphology of neurons was observed with optical microscope. The expression of autophagy-related protein (LC3, P62) and apoptosis-related protein (cleaved caspase-3) were assessed by Western Blot analysis. The apoptosis of neurons was detected by flow cytometry, the release rate of lactate dehydrogenase (LDH) was calculated by automatic biochemical analyzer, and the cell activity was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay.Results Compared with the control group, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.091±0.160, 3.422±0.186 vs. 0.256±0.021), cleaved caspase-3 (gray value: 0.230±0.025, 0.440±0.051 vs. 0.050±0.007), neuronal apoptotic rate, LDH release rate [(38.50±4.15)%, (59.60±5.65)% vs. (12.40±1.32)%] were increased, while the expression of P62 (gray value: 0.290±0.025,0.120±0.026 vs. 0.450±0.040), neuronal activity [(71.40±7.23)%, (42.80±4.12)% vs. (100.30±2.30)%] were decreased at 2 hours or 6 hours after OGD (allP < 0.05). When the time of OGD was 2 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ (gray value: 2.281±0.121), the neuronal activity [(51.10±5.73)%] were decreased, while the expression of P62 and cleaved caspase-3 (gray scale: 0.410±0.037, 0.330±0.027, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (47.30±4.43)%] were increased (allP < 0.05). When the time of OGD was 2 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ (gray value: 3.689±0.214), the neuronal activity [(85.30±8.56)%] were increased, while the expression of P62 and cleaved caspase-3 (gray value: 0.170±0.040, 0.090±0.096, respectively), neuronal apoptotic rate, the injury of neurons [LDH release rate: (24.30±2.14)%] were decreased (allP < 0.05). On the contrary, when the time of OGD was 6 hours and it was combined with 3-MA, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.021±0.178, 0.240±0.017), neuronal apoptotic rate, the injury of neurons [LDH release rate: (36.60±3.45)%] were decreased, while the expression of P62 (gray value: 0.350±0.060), the neuronal activity [(59.70±6.13)%] were increased (allP < 0.05). When the time of OGD was 6 hours and it was combined with rapamycin, the expression of LC3 Ⅱ/Ⅰ and cleaved caspase-3 (gray value: 3.923±0.201, 0.590±0.062), neuronal apoptotic rate, the injury of neurons [LDH release rate:(71.20±7.81)%] were increased, while the expression of P62 (gray value: 0.070±0.008), the neuronal activity [(27.30±2.12)%] were decreased (allP < 0.05).Conclusion The enhancement of autophagy has protective effect on neurons under the condition of mild OGD, while it can aggravate the injury of neurons induced by a long-time OGD.

Adult ; Comet Assay ; DNA Damage ; drug effects ; Female ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Peracetic Acid ; toxicity