Objective To test the effects of restuscitation with air or oxygen on the blood gas and cerebral superoxide dismutase (SOD) concentration in neonatal rats with experimental intrauterine asphyxia. Method Seventy-seven neonatal rats were randomly (random number) divided into three experimental groups: sham operation group (SHAM), air resuscitation group (AR), and oxygen resuscitation group (OR). In groups AR and OR, animal models of intrauterine asphyxia were established and then resuscitated with air (AR) or oxygen (OR) for 30min. Comparison was made between groups including the mortality 0 hour after resuscitation, and the levels of blood gas and cerebral SOD concentrations 0 h, 6 h and 24 h after resuscitation. Results Mortality of neonatal rats in SHAM group, AR group and OR group were 0 (0/24), 0 (0/26) and 3.7% (1/27), respectively (P ＞0.05). The average levels of blood PaO2 in OR group and AR group 0 h after resuscitation were (69.2 ± 8.2)mmHg and (55.5±10.3) mmHg,respectively (P=0.004). Blood pH and PaCO2 and BE levels in OR group 0 h after resuscitation were not significantly different from those in AR group (P＞0.05). Blood pH, PO2, PCO2and BE levels in OR group were also not significantly different from those in AR group 6 h and 24 hours after resuscitation. The average concentrations of cerebral SOD in OR group 0 h and 6 hours after resucitation were (38.3±9.8) U/mgprot and (8.6±3.6) U/mgprot, and those in AR group were (53.8± 10.6) U/mgprot and (13.0±4.6) U/mgprot, respectively (P = 0.003, 0.04). The cerebral SOD concentration in OR group 24 hours after resuscitation was not significantly different from that in AR group (P＞0.05). The cerebral SOD concentrations in SHAM group 0 h,6 h and 24 hours after resuscitation were much higher than those in OR group and AR group (P＜0.05). Conclusions Resuscitation with air is as good as pure oxygen in neonatal resuscitation, in respect of early mortality and improvement of acidosis in neonatal rats after intrauterine asphyxia. Resuscitation with air will generate less radical oxygen species than pure oxygen in neonatal rats after intrauterine asphyxia.

Objective To study the relationship between C reactive protein ( CRP) and postoperative deliri-um of elderly patients after hip fracture , and to explore the possible mechanism of delirium .Methods A total of 186 elderly patients with hip fracture who received operation were included in the study .24h after operation,according to the patients'awareness of ICU fuzzy evaluation method ,the patients were divided into the delirium group and non delirium group .The levels of CRP and procalcitonin were detected and compared .Results The CRP levels in the de-lirium group and non delirium group were (207 ±86)mg/L,(87 ±42)mg/L,there was significant difference between two groups (t =2.427,P <0.01).The procalcitonin levels in the delirium group and non delirium group were (0.32 ±0.11)μg/L,(0.27 ±0.13)μg/L,the difference between the two groups was not significant (t=0.347,P>0.05).Conclusion CRP may be related with delirium.Inflammatory response without systemic infection may be one of the mechanisms of delirium .

Objective To study effect of Xuebijing injection on coagulation function and prevention of deep venous thrombosis in elderly fracture patients with mechanical ventilation.Methods 60 patients were randomly divided into control group(30 cases) and Xuebijing group(30 cases).Xuebijing group received Xuebijing injection, besides physical therapy for prevention of deep venous thrombosis received by control group.Coagulation parameters and Lac at different time points,thrombosis incidence,hemorrhage incidence APACHE II score and 28 -day mortality were compared between the two groups.Results In Xuebijing group,PT,APTT,DD on d3 and Fg 48 on d5 had statistically significant differences compared with before treatment.PT,APTT on d3,d5 and DD on d5 were statistical-ly different compared with the control group(P <0.05).PLT,Lac,hemorrhage incidence and 28 -day mortality had no significant differences between the two groups.Thrombosis incidence in Xuebijing group was significantly lower than the control group(20.00% vs 3.33%) (χ2 =4.043,P =0.044,P <0.05).Both of APACHE II score were lower than before treatment,but the Xuebijing group was significantly decreased(t =5.48,P =0.000,P <0.05). Conclusion Xuebijing injection could improve coagulation function,and decrease thrombosis incidence in elderly fracture patients with mechanical ventilation.

Objective To construct a new recombinant immunotoxin expression vector fused with a murine interleukin18(IL18) gene and a truncated pseudomonas exotoxin (PE38) gene, and examine the expression of IL-18-PE38 fusion protein in Escherichia coli (E. coli). Method Murine IL-18 (mIL-18) cDNA was cloned from murine liver tissue through reverse transcription-polymerase chain reaction (RT-PCR). The mIL-18 cDNA was ligased with a PE38 gene carried by PRKL expression vector through T4 DNA ligase and constructed into fusion protein expression plasmid PRKL-IL18-PE38. The recombinant vector was identified by restriction endonucleases digestion, PCR and DNA sequencing. After transformed into E.coli BL21 and induced by IPTG, the expressed product was obtained and the molecular weight and specificity were determined by SDS-PAGE and Western-blotting. Result The new recombinant immunotoxin expression vector was constructed successfully. DNA sequencing revealed that the mIL-18 and PE38 gene were consistent with NCBI Gene Bank. The IL-18-PE38 fusion protein was expressed in E.coli BL21, and Western-blotting analysis indicated that the molecular weight of the expression product is about 56 kDa, and could react with the specific antibody against mIL-18. Conclusion IL-18-PE38 recombinant immunotoxin expression vector will provide the basis for study on the targeted cytotoxic activity to Th1 cells and may have some potential value in the treatment of Th1 cell-mediated autoimmune diseases.

Objective To discuss the clinical application of homemade airway exchange catheter on the extubation of patients with difficult tracheal intubation in intensive care unit(ICU). Methods Sixty-two patients with difficult tracheal intubation who failed their initial extubation trial were randomly divided into conventional group (31 cases)and observation group(31 cases). The patients in the conventional group received routine extubation process,while the patients in the observation group were extubated under the guidance over a homemade airway exchange catheter. The changes in heart rate(HR),blood pressure,respiratory rate(RR)and pulse blood oxygen saturation(SpO2)were compared at 12 hours after extubation,so as the re-intubation rate,intubation success rate at first attempt and re-intubation time in two groups,and the tolerance and complications after extubation were observed. Results After extubation,the HR,blood pressure and RR were increased significantly(all P<0.05), and the SpO2 was much lower in conventional group(P<0.05),while those parameters were changed little and basically in the normal ranges in the observation group. At 12 hours after extubation,the re-intubation rate was much lower(6.45%vs. 25.81%,P<0.05)in the observation group,with shorter re-intubation time(seconds:27±14 vs. 49±28,P<0.01),higher intubation success rate at first attempt(90.32%vs. 54.84%,P<0.01)and better tolerance (77.4% vs. 61.3%,P<0.05)compared with those in the conventional group. There was no severe complication in the observation group,and there were 1 cases of glottic edema with cricothyroid membrane puncture,2 cases of broncheal mucous membrane bleeding and 2 cases of bucking in the conventional group. Conclusion Compared with conventional extubation process,the extubation over homemade airway exchange catheter can increase the rate of extubation,reduce re-intubation rate and the re-intubation time,with favorable tolerance and no occurrence of serious complications,and is one of the safe and effective extubation strategies in patients with difficult tracheal intubation in ICU.

In order to explore the feasibility and protective efficiency of influenza DNA vaccine, we constructed eukaryotic expressing plasmids encoding HA and HA1 of influenza A virus (A/PR/8/34) and studied their expression in HEK293 cells. HA and HA1 genes were amplified by RT-PCR and cloned into pcDNA3.1(+) to generate pcDNA3.1(+)/HA and pcDNA3.1(+)/HA1, respectively. After verification of the cloning fidelity by restriction endonuclease digestion, PCR, and sequencing, pcDNA3.1(+)/HA and pcDNA3.1(+)/HA1 were transfected into HEK293 cells using PolyFect Transfection Reagent. Immunofluorescence assay was used to detect the transient expressing cells. Fluorescence microscopy revealed strong expression of target gene in HEK293 cells transiently transfected with either pcDNA3.1(+)/HA or pcDNA3.1(+)/HA1. Therefore, the results confirm the successful construction of eukaryotic expressing plasmids capable of driving the eukaryotic expression of influenza virus antigen HA and HA1, which is likely to provide a basis for both further investigation of the mechanism of influenza viral infection and the development of influenza DNA vaccine.

Objective To study the effect of Xuebijing injection on transfusion and systemic inflammation in traumatic coagulopathy patients. Methods Seventy-nine patients were randomly divided into Xuebijing Group (36 cases) and Control Group (43 cases). Xuebijing group received Xuebijing injection in addition to the regular treatment received by control group. Levels of TNF-α, IL-6, IL-8 and transfusion amount between two groups were compared. Results Coagulation abnormality was almost corrected in all patients after treatment. In Xuebijing Group, transfusion of RBC, FFP and cryoprecipitate were significantly less than that in control group (P<0.05). At 72 h after admission, TNFα, IL-6 and IL-8 were significantly lower in Xuebijing Group (P < 0.05). Conclusion Xuebijing injection could reduce transfusion in traumatic coagulopathy patients , possibly resulting from its antagonism against systemic inflammation.

Objective: To investigate the immunological mechanism of influenza A virus for murine S180 ascites sarcoma. Methods: After inoculation with S180 sarcoma cells, mice were i. p. injected with influenza A virus or vehicle 15 days. The average living time and survival rate of the mice were examined. The levels of IL-2, IL-6 and TNF-? were detected. The sarcoma cell's apoptosis was detected by DNA ladder, flow cytometry (FMC) , fluorescent microscope and e-lectron microscope (EM). Results: The average living time and survival rate of the mice injected with Influenza A virus were significantly longer or higher than that of the controls. The levels of IL-2, IL-6 and TNF-? also had the same differences. The apoptosis cells were detected by EM and fluorescent microscope. Sub-diploid peaks were observed by FCM a-nalysis and DNA ladder was seen after electrophoresis in the ascites cells. Conclusion: Our results demonstrated that the feasibility and potential of delivery of influenza A virus as a general means for the treatment of S180 ascites sarcoma.

Objective To investigate the relationship between RELN gene single nucleotide polymorphism (SNP) and childhood autism in Jiamusi, Heilongjiang. Methods Polymerase chain reaction-restricted fragment length polymorphism (PCR-RFLP) was used to determine allele and genotype of SNP (exon 6) of RELN in 30 children with autism and 30 normal children. Autism Behavior Checklist (ABC) was used to evaluate the children. Results There was a significant difference in the distribution of the allelic frequencies and genotype in exon 6 between these groups (P<0.05). There was a significant difference in the communication factors between patients with genotype of A/A and A/G or A/A and G/G (P<0.05), as well as in the total scores of ABC between A/G and G/G (P<0.05). Conclusion The SNP of RELN (exon 6) associated with the childhood autism. There is a more serious communication disorder in children with genotype of G/G, A/G than that of A/A.

In order to explore the feasibility and protective efficiency of influenza DNA vaccine, we constructed eukaryotic expressing plasmids encoding HA and HA1 of influenza A virus (A/PR/8/34) and studied their expression in HEK293 cells. HA and HA1 genes were amplified by RT-PCR and cloned into pcDNA3. 1 (+) to generate pcDNA3. 1 (+)/HA and pcDNA3.1 (+)/HA1, respectively. After verification of the cloning fidelity by restriction endonuclease digestion, PCR, and sequencing, pcDNA3.1 (+)/HA and pcDNA3.1 (+)/HA1 were transfected into HEK293 cells using PolyFect Transfection Reagent. Immunofluorescence assay was used to detect the transient expressing cells. Fluorescence microscopy revealed strong expression of target gene in HEK293 cells transiently transfected with either pcDNA3. 1 (+)/HA or pcDNA3. 1 (+)/HA1. Therefore, the results confirm the successful construction of eukaryotic expressing plasmids capable of driving the eukaryotic expression of influenza virus antigen HA and HA1, which is likely to provide a basis for both further investigation of the mechanism of influenza viral infection and the development of influenza DNA vaccine.