OBJECTIVE To investigate the ?lactamase genes in Escherichia coli and their resistance.METHODS Twenty strains of E.coli producing extended spectrum ?-lactamases(ESBLs) were isolated from hospitalized patients.?-Lactamase drug-resistant genes were detected by PCR.RESULTS There were higher positive rates of ?-lactamase genes of TEM,SHV,LEN,CTX-M-1,CTX-M-9,VEB,GES,PER(75.0%,20.0%,95.0%,15.0%,70.0%,5.0%,85.0% and 70.0%,respectively). This study showed that there were high positive rate of drug resistance.CONCLUSIONS There are very high positive percentages of TEM,LEN,CTX-M-9,GES,and PER genes in ESBLs producing E.coli isolated clinically,and showing highly drug-resistant E.coli of producing ESBLs can be transmitted in hospital and induce fulminant prevalence.

Objective To investigate the effects of adenosine postconditioning (AP) on serum IL-10 and TNF-α concentrations following myocardial ischemia-reperfusion(VR)in rats.Methods Twenty-four SD ratsweighing 180-250 g were randomly divided into 4 groups(n=6 each):group I sham operation (group S);group Ⅱ myocardial I/R;group Ⅲ ischemic postconditioning(group IP)and group Ⅳ AP.Myocardial I/R was induced by 30 rain occlusion of anterior descending branch of left coronary artery followed by 120 min reperfnsion.IP was induced by 3 cycles of 30 s myocardial ischemia followed by 30 s reperfusion at the end of ischemia.In AP group adenosine 1.5 mg/kg was infused at 40μg·kg-1·min-1 before the onset of reperfusion.SP,DP and HR were recorded before ischemia (baseline) at 30 min of ischemia and 30 and 120 min of reperfusion.Arterial bloodsarnples were collected at 120 min of repednsion for determination of serum TNF-α and IL-10 concentrations.Theanimals were then killed.Their hearts were removed for microscopic examination.Myocardial infarct size wasmeasured and myocardial MDA content was determined.Results BP and HR were signilicandy decreased duringreperfusion while myocardial infarct size.MDA content and serum concentrations of IL-10 and TNF-α weresignificantly increased in I/R group compared with group S.Ischemic and adenosine postconditioning significantlyattenuated hypotension,reduced infarct size,myocardial MDA content and serum TNF-α concentration and increased serum IL-10 concentration in group AP and IP as compared with I/R group.There was no significant difference in the above changes between group AP and IP. Myocardial injury was ameliorated in group AP and IP as compared with I/R group. Conclusion Adenosine postconditioning can protect myocardium from I/R injury by increasing IL-10 production and inhibiting TNF-a release.

Objective To choose zebrafish as the experimental animal model for studying the spatiotemporal expression rule of rap1 gen in zebrafish embryo early development process .Methods The Rap1 gene fragment was cloned from the zebrafish emby‐oscDNA ,then the Rap1 gene fragment and pCS2+ plasmid were performed the in vitro connetion and recombination was extracted , the combinant plasmid was correct after the double enzyme digestion ,colony PCR and sequencing identification .T3 RNA polymer‐ase in vitro transcription system was used to obtain the digoxin (DIG)‐labeled anti‐sense mRNA probe of Rap1 gene .The whole mount in situ hybridization method was adopted to detect the Rap1 expression in zebrafish embryo early development process . Results The positive hybridization signal of Rap1 gene was detected at the cell division junction region of 0 .75 hpf ,animal pole of 3 .70 hpf and 6 .00 hpf ,and notochord of 12 .00 -72 .00 hpf .Conclusion Rap1 gene might be involved in the early development process of notochord nervous system in zebrafish .

OBJECTIVETo evaluate the curative effect of external lumbar drainage combined with empirical antibiotics treatment on the postoperative culture negative meningitis.

METHODSThe clinical data of eighty post-operative meningitis patients with cerebrospinal fluid culture negative were retrospectively analyzed according to inclusive and exclusive criteria from January 2013 to December 2014 in Department of Neurosurgery, Tianjin Huanhu Hospital. All patients were composed of 45 male cases and 35 female cases, aging from 9 to 72 years. All patients were divided into two groups according to receiving the different treatment: one group only receiving a intravenously empirical antibiotics treatment (n=40), another group receiving a combined therapy of external lumbar drainage and intravenously empirical antibiotics treatment (n=40). The volume of drainage of cerebrospinal fluid (CSF) were set up from 200 to 300 ml per day. There was no difference in the dosage and interval of the same antibiotics between two groups. The antibiotics usage and therapeutic effect of two groups of patients were observed and analyzed by t-test, Wilcoxon rank test or χ(2) test.

RESULTSThe rate of CSF bacterial culture negative was 62.9% (88/140) in the same period. In group of empirical antibiotics treatment, the time of antibiotics treatment was (12.6±3.1) days, the rate of combined with other antibiotics treatment was 40.0% (16/40), the rate of mortality was 15.0% (6/40). However, in group of external lumbar drainage combined with empirical antibiotics treatment, the 3 data were (5.3±1.2) days, 10.0% (4/40), 7.5% (3/40), respectively. The time of antibiotics of the group of empirical antibiotics was longer (t=3.605, P=0.017), while the rate of combined antibiotics and the rate of mortality were lower (χ(2)=3.971, P=0.035; χ(2)=4.136, P=0.027, respectively). The average drainage time was (5.8±1.5) days, 32 patients gained a complete healing only by their first placement, 5 cases need replacement because of occlusion or drainage time more than 14 days. There were no recurrence cases after 3 months' follow-up.

CONCLUSIONSThe method of external lumbar drainage combined with empirical antibiotics treatment can significantly reduce the dosage and course of antibiotics treatment. It is a simple, safe, efficacious method for the treatment of neurosurgical postoperative bacterial culture negative meningitis.

Adolescent ; Adult ; Aged ; Anti-Bacterial Agents ; therapeutic use ; Child ; Drainage ; Female ; Humans ; Male ; Meningitis ; drug therapy ; therapy ; Middle Aged ; Neurosurgical Procedures ; Postoperative Complications ; drug therapy ; Retrospective Studies ; Young Adult

Objective To evaluate the effect of quercetin pretreatment on the permeability of blood-brain barrier in a rat model of global cerebral ischemia-reperfusion ( I∕R). Methods Sixty-three clean-grade healthy male Sprague-Dawley rats, weighing 300-350 g, aged 4-5 months, were divided into 3 groups (n＝21 each) using a random number table method: sham operation group ( group S), group I∕R and quercetin pretreatment group ( group Q). Global cerebral I∕R was induced by occlusion of bilateral common carotid arteries combined with hypotension ( mean arterial pressure was maintained at 35-45 mmHg) in chloral hydrate-anesthetized rats. Quercetin 25 μmol∕kg was injected intraperitoneally twice a day for 3 consecutive days starting from 3 days before establishment of the model in group Q, while the e-qual volume of normal saline was given instead at the corresponding time points in group S and group I∕R, respectively. The animals were sacrificed at 24 h of reperfusion and brains were removed to determine the brain water content, Evans blue ( EB) content and expression of occludin protein in cerebral cortex ( by Western blot) and to observe the ultrastructure of blood-brain barrier. Results Compared with group S, the brain water content and EB content were significantly increased, the expression of occludin protein was down-regulated (P＜0. 05), and the injury to ultrastructure of blood-brain barrier was accentuated in I∕R and Q groups. Compared with group I∕R, the brain water content and EB content were significantly de-creased, the expression of occludin protein was up-regulated (P＜0. 05), and the injury to ultrastructure of blood-brain barrier was significantly attenuated in group Q. Conclusion Quercetin pretreatment can de-crease the permeability of blood-brain barrier and attenuate brain edema, and the mechanism may be related to up-regulated expression of occludin protein in a rat model of global cerebral I∕R.

Objective To evaluate the relationship between the pathomechanism of neuropathic pain (NP)-inducced depression and autophagy in the cortex of the frontal lobe in rats.Methods Forty healthy male Sprague-Dawley rats in which IT catheters were successfully placed,aged 8-10 weeks,weighing 200-220 g,were divided into 4 groups (n =10 each) using a random number table method:sham operation group (group S),group NP,NP plus dimethyl sulfoxide group (group ND) and NP plus autophagy inducer rapamycin group (group NR).The neuropathic pain model was established by ligation of the left fifth spinal nerve of anesthetized rats in NP,ND and NR groups.Rapamycin 0.1 μgwas intrathecally injected via the intrathecal catheter immediately after ligation of the spinal nerve and every day after ligation once a day for 21 consecutive days in group NR.The equal volume of dimethyl sulfoxide was intrathecally injected instead of rapamycin in group ND.The mechanical paw withdrawal threshold (MWT) was measured before ligation and at 1,3,7,10,14 and 21 days after ligation.The forced swimming test was performed at 3 days before ligation and 14 and 21 days after ligation.The rats were sacrificed after the last measurement of the behaviour testing,and the prefrontal cortex was removed for determination of the expression of microtubule-associated protein light chain 3 Ⅰ (LC3 Ⅰ) and LC3 Ⅱ,Beclin-1 and p62 (by Western blot).The ratio of LC3 Ⅱ/LC3 Ⅰ was calculated.Results Compared with group S,the MWT was significantly decreased after ligation,the time of immobility was prolonged,the expression of LC3 Ⅰ was down-regulated,the expression of LC3 Ⅱ,Beclin-1 and p62 was up-regulated,and the LC3 Ⅱ/LC3 Ⅰ ratio was increased in NP and ND groups (P＜0.05).Compared with group NP,the MWT was significantly increased after ligation,the time of immobility was shortened,the expression of LC3 [and p62 was down-regulated,the expression of LC3 Ⅱ and Beclin-1 was up-regulated,and the LC3 Ⅱ/LC3 Ⅰ ratio was increased in group NR (P＜0.05).Conclusion Enhanced autophagy in the cortex of the frontal lobe is involved in the endogenous antidepressant mechanism in rats with NP.

Objective To evaluate the advantages of sevoflurane inhalation anesthesia combined with lumbosacral plexus block for hip replacement in elderly patients. Methods Sixty patients of both se-xes, aged 65-85 yr, with body mass index of 18-25 kg∕m2, of American Society of Anesthesiologists physical statusⅠ-Ⅲ, scheduled for elective unilateral hip replacement, were divided into 2 groups(n=30 each)using a random number table: total intravenous anesthesia group(TIVA group)and GasMan software-guided sevoflurane inhalation anesthesia combined with lumbosacral plexus block group(SEV-B group). After the laryngeal mask airway was inserted, the patients were mechanically ventilated in both groups. Heart rate and mean arterial pressure(MAP)were recorded before anesthesia(T0), immediate-ly after insertion of laryngeal mask airway(T1), immediately after skin incision(T2), immediately after intramedullary reaming(T3), immediately after prosthesis implanting(T4), at the end of surgery(T5) and at 5 min after removal of the laryngeal mask airway(T6). The end-tidal concentration of sevoflurane at T2-5was also recorded in group SEV-B. The time for removal of the laryngeal mask airway, duration of stay in post-anesthesia care unit and first ambulation time were recorded. The consumption of anesthetics used during patient-controlled intravenous analgesia(PCIA), effective pressing times of PCIA, requirement for rescue analgesic and adverse reactions were also recorded at 24 h after surgery. The patient′s cognitive func-tion was assessed using Mini-Mental State Examination at 1 day before surgery and 1 and 3 days after surger-y, and the occurrence of postoperative cognitive dysfunction was recorded. Results Compared with group TIVA, heart rate at T1-6and MAP at T2-6were significantly decreased, and MAP at T1was increased in group SEV-B, and the time for removal of the laryngeal mask airway, duration of stay in post-anesthesia care unit and first ambulation time were significantly shortened, the consumption of anesthetics used during PCIA, effective pressing times of PCIA, requirement for rescue analgesic and incidence of adverse reac-tions were reduced at 24 h after surgery, the Mini-Mental State Examination scores were increased at 1 day after surgery, and the incidence of postoperative cognitive dysfunction was decreased at 1 day after surgery in group SEV-B(P<005). Conclusion GasMan software-guided sevoflurane inhalation anesthesia com-bined with lumbosacral plexus block can provide more accurate anesthesia management for hip replacement and promote rapid rehabilitation after surgery in elderly patients.

Objective To evaluate the effect of continuous low-to medium-flow oxygen administration in non-ventilated lung on the oxidative stress response of lung tissues during one-lung ventilation (OLV).Methods Fifty-seven American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 50-64 yr,weighing 40-74 kg,scheduled for elective pulmonary tumorectomy,were divided into 2 groups using a random number table:control group (group C,n=29) and continuous oxygen administration group (group O,n =28).The patients were intubated with the double-lumen tube after induction of anesthesia.Correct position of the tube was verified with the fiberoptic bronchoscope.In group O,the F14 tube was placed at 2-3 cm beyond the carina of trachea in the non-ventilated lung at the beginning of OLV,and low-to medium-flow oxygen was continuously administered at 1-4 L/min with the fractional concentration of inspired oxygen set at 25％-37％.Blood samples were taken from the radial artery and internal jugular bulb at the beginning of anesthesia induction (T1) and 30 min,1 h and 2 h of OLV (T2-4) for blood gas analysis.Lung tissues at the site 5 cm lateral to the tumor were taken immediately after resection of diseased tissues for determination of superoxide dismutase and malondialdehyde levels (by chemical colorimetry) and heme oxygenase-1 expression (by Western blot).Results Compared with group C,the partial pressure of arterial oxygen was significantly increased at T2-4,the partial pressure of arterial carbon dioxide was decreased at T2,3,the partial pressure of venous oxygen was increased at T2,3,the partial pressure of venous carbon dioxide was decreased at T2-4,the malondialdehyde level was decreased,and the expression of heme oxygenase-1 was up-regulated (P＜0.05),and no significant change was found in superoxide dismutase level in group O (P＞0.05).Conclusion The mechanism by which continuous low-to medium-flow oxygen administration in non-ventilated lung exerts pulmonary protection is related to inhibiting oxidative stress responses of lung tissues during OLV.

Objective: To evaluate the effect of irisin preconditioning on global cerebral ischemia-reperfusion (I/R) injury in rats. Methods: Thirty-six healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 3 groups (n=12 each) using a random number table method: sham operation group (group S), global cerebral I/R group (group I/R) and irisin preconditioning group (group I). Global cerebral I/R was induced by occlusion of bilateral common carotid arteries combined with hypotension (MAP maintained at 35-45 mmHg) in anesthetized rats.At 30 min before ischemia, irisin 10 μg/kg (diluted to 10 μg/ml in normal saline) was intravenously injected in group I, and the equal volume of normal saline was intravenously injected in S and I/R groups.Morris water maze test was performed at day 3 of reperfusion to assess the cognitive function.Rats were sacrificed after the end of morris water maze test, and brains were removed for determination of histopathologic changes in hippocampal CA1 region (using HE staining), neuronal apoptosis in hippocampal CA1 region (Tunel staining), glial fibrillary acidic protein (GFAP) expression (by Western blot), myeloperoxidase (MPO) activity in the hippocampal tissues (by colorimetric assay), and contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in hippocampal tissues (by enzyme-linked immunosorbent assay). The cell necrosis rate and apoptotic rate were calculated. Results: Compared with group S, the escape latency was significantly prolonged on 1-5 days in group I/R and on 1-3 days in group I, the time of staying at 1st quadrant was significantly shortened, the cell necrosis rate and apoptotic rate were increased, the expression of GFAP was up-regulated, and the activity of MPO and contents of TNF-α and IL-1β were increased in I/R and I groups (P<0.05 or 0.01). Compared with group I/R, the escape latency was significantly shortened on 1-5 days, the time of staying at 1st quadrant was prolonged, the cell necrosis rate and apoptotic rate were decreased, the expression of GFAP was down-regulated, and the MPO activity and contents of TNF-α and IL-1β were decreased in group I (P<0.05 or 0.01). Conclusion Irisin preconditioning can reduce the global cerebral I/R injury in rats, and the mechanism may be related to inhibiting activation of astrocytes in hippocampus and reducing inflammatory responses.