Objective To investigate the value of salivagram in the diagnosis of pediatric pulmonary aspiration.Methods A total of 469 pediatric patients (age:6 weeks-16 years,average age (4.5±3.1) years;229 males,240 females) underwent salivagram.Signed informed consents were obtained.99Tcm-SC (11.1 MBq,100 μl) was dropped into the root of tongue.After the dropping,the posterior dynamic acquisition was started for 60 min,and then the anterior and posterior static images were acquired for 5 min.All patients were followed up for 3-6 months after the salivagram,and the salivagram results were analyzed.If radioactivity was shown in the main bronchus or bilateral lung fields,the pulmonary aspiration was diagnosed.Results Salivagram demonstrated positive results in 20.7％ (97/469) of the patients.However,3 patients with pulmonary aspiration showed negative results.There was no false positive result on salivagram.Conclusion Salivagram shows good results in the diagnosis of pediatric pulmonary aspiration.

Objective Making the fusion protein of IgG-binding peptide with enhanced green fluorescent protein(EGFP) and determining its bioactivity.Methods The enhanced green fluorescent protein(EGFP) gene was cloned into pEZZ 18 vector containing ZZ peptide gene to construct expression vector pSpA-EGFP-His.The fusion protein was expressed in E.coliDH5? and its bioactivity was detected by competitive ELISA and fluorescence properties.Results The fusion protein migrated at approximately 42kD in SDS-PAGE,which correspond to the theoretical molecular weight.The spectra of SpA-EGFP fusion protein was similar to what was reported.SpA-EGFP competed with SpA-Peroxidase to bind IgG.Conclusion The plasmid pSpA-EGFP-His correctly expressed in E.coli.The fusion protein retains the bifunctional effects of EGFP and IgG-binding activity.

Objectives:To construct rGA antisense eukaryotic expression vector. Methods:Using molecular cloning technology, we subcloned rGA cDNA in its antisense orientations of the pcDNA 3.0 vector. Results:Cutting with Eco RⅠ and Hin D Ⅲ,antisense recombinant plasmid produce 2 bands. One was 0.28 kb fragment, the other was 5.5 kb fragment which corresponded to theoretical calculation. Conclusions:We succeeded in constructing rGA antisense eukaryotic expression vector.

Objective To observe the variation of plasma 8-hydroxy-2'-deoxyguanosine (8-OHdG),a biomarker of oxidative DNA damage,in patients with Alzheimer's disease (AD) versus vascular dementia (VaD),and to investigate the important role and the difference of oxidative DNA damage in the initiation and development of AD and VaD.Methods 35 AD patients,28 VaD patients and 23 age and sex matched normal controls were enrolled in this study.Plasma 8-OHdG was measured by an enzyme-linked immunosorbent assay (ELISA) method.Cognition function in AD and VaD patients was measured by Mini-Mental State Examination (MMSE).Results There were significant differences in plasma 8-OHdG concentrations among AD,VaD and control groups [(601.8±266.0) ng/L,(1748.0±665.6)ng/L,(352.0±94.5)ng/L,respectively,P＜0.001].Plasma 8-OHdG was significantly higher in VaD group than in control group (P＜0.05).While,there was no significant difference in plasma 8OHdG concentration between AD group and control group (P＞0.05).Plasma 8-OHdG was significantly higher in VaD group than in AD group (P＜0.05).Plasma 8-OHdG levels were positively related with age in AD,VaD and control groups (r=0.661,0.702,0.719,respectively,all P％0.01).No correlations were found between plasma 8-OHdG concentration and education level among the three groups (all P＞0.05).AD and VaD patients were subgrouped according to the risk factors including gender,smoking,hypertension,diabetes,hyperlipidemia.There were no significant differences in plasma 8-OHdG level between the subgroups (all P＞ 0.05).After adjusting for sex,age,education level,smoking,hypertension,hyperlipidemia and diabetes,logistic regression analysis showed that the probability of high plasma 8-OHdG level was higher in VaD group than in AD and control groups.Plasma 8-OHdG level had a negative correlation with MMSE score in VaD group (r=-0.592,P＜0.01),while no correlation was found between plasma 8-OHdG level and MMSE score in AD group (r =-0.122,P＞ 0.05).Conclusions There is significant oxidative DNA damage in VaD patients.Plasma oxidative DNA damage is more serious in VaD patients than in AD patients and healthy people.Plasma 8-OHdG level,a oxidative DNA damage biomarker,has a significant correlation with the degree of cognition impairment in VaD patients.

Objective To construct the recombinant plasmid carrying antisense GA cDNA, and evaluate the effect of antisense GA gene on the biological characteristics of colon cancer cells. Methods The recombinant plasmid pcDNA3.0/GA containing antisense GA gene was constructed by gene recombination technique. The recombinant plasmid was transfected into colon cancer cells by lipofectamine. The effect of antisene GA gene on the biological characteristics of colon cancer cell line DH52 was evaluated by MTT and electron microscopy. Results The recombinant plasmid pcDNA3.0/GA was successfully constructed. The growth rate of colon cancer cells decreased, and microstructure of the cells appeared typical apoptotic changes after transfection. Conclusion Antisense GA gene could inhibit the growth of colon cancer cells possibly by suppressing GA gene expression and decreasing GA activity in the colon cancer cells.

Objective To investigate whether intestinal bacterial translocation in scalded rat with delayed fluid resuscitation (DFR) was partly due to the reduction of the expression of rat defensin (RD), which was the most important defensin in rat intestinal defensin family excreted by Paneth cells. Methods Fifty-six SPF rats were randomized to 3 groups: sham injury group (n=8); early resuscitation (ER) group (n=24), receiving fluid resuscitation immediately after scald (30% TBSA, third degree); delayed resuscitation group (n=24), receiving fluid resuscitation 6 hours after scald. The animals (n=6, at each time point) were sacrificed at 8, 24, 72 hours after injury. The expression of RD-5 in the terminal ileum was determined with PCR technic. Morphological changes in ileal Paneth cells were observed. Quantitative bacterial cultures of mesenteric lymph nodes (MLN), liver, spleen and lung were also done. Results In both ER group and DR group, ileal RD-5 mRNA expression level was significantly increased at 8 hours after injury, and it began to decrease at 24 hours until the end of the experiment, when the level was much lower than that of the sham group. When two experimental groups were compared, RD-5 mRNA expression level was significantly higher at 8 hours and significantly lower at 72 hours in DR group than in ER group. There were no obvious morphological changes in ileal Paneth cells in all 3 groups. Although high incidence of bacterial translocation was observed in both experimental groups, it was significantly higher in DR group than in ER group. Conclusions DFR did not cause Panth cell damage, and RD-5 changes did not contribute to bacterial translocation in 24 hours after DFR, as the increase in RD-5 mRNA might be an innate protective reaction. However, the decrease in RD-5 production at 72 hours may play a role in bacterial transloca- tion.

Objective To explore the diagnosis and topical treatment of diabetic foot in patients with burns.Methods Sixty-five patients with burns complicated with diabetic foot were admitted from Jan.2002 to Dec.2006.Bacterial culture of wound excretion,peripheral nerve electromyography and vascular ultrasonography of both lower extremities were performed in a part of patients.Twenty-four patients received topical treatment,including immersion bath with hypertonic saline solution and the application of epidermal growth factor,fibroblast growth factor and complex lysozyme disinfectant.Forty-one patients underwent surgical treatment.The operation methods,surgical treatment included debridement followed by direct closure,split-thickness skin grafting,repair with local skin flap or cross leg skin flap,or amputation of foot,as determined according to the condition of the wound.Results The positive rate of bacterial culture of wound excretion was 88.7%(47/53)on admission and 30.2%(16/53)one week after topical treatment.There were abnormal findings of peripheral nerve electromyography in all 31 cases.Vascular ultrasonography revealed pathological findings in both lower extremities of all 12 patients with vascular lesion,and 9 in 16 cases without vascular lesion.The lesions in all patients except 2,who were discharged in advance,healed and the cure rate was 96.9%.Conclusion Peripheral nerve electromyography and vascular ultrasonography of both lower extremities are promising auxiliary examinations for the diagnosis of diabetic foot in patients with burns.Topical treatment,which includes immersion bath with hypertonic saline solution and the application of growth factors and complex lysozyme disinfectant,as well as appropriate surgical management,plays an important role in the wound healing.

Objective To observe the effect of low molecular weight heparin(LMWH)on hypercoagulability in extensive severe burn patients.Methods 9 patients were assigned as LMWH treatment group(TG),and another 12 burn patients with the similar extent of burn injury admitted in the same period were assigned as routine treatment group(RTG).All the treatments were the same except 5000U LMWH was given via subcutaneous injection per 12 or 6 hours to the TG group for 3-7days.The patients in normal control group(NCG)consisted of patients receiving plastic surgery.The following coagulation parameters were determined before and after the heparin treatment:prothrombin time(PT),thrombin time(TT),activated partial thromboplastin time(APTT),international normalized ratio(INR),fibrinogen(Fib)and platelet(PLT).Results INR,APTT and PLT in both TG and RTG groups were significantly lower compared with that in NCG before treatment,and Fib in two former groups were higher than that in NCG.Compared with those values before treatment,Fib decreased and PLT increased significantly after LMWH treatment in TG.However,there were no significant changes in other parameters in TG after LMWH treatment.Severe side effects such as haemorrhage were not found in patients in TG.Conclusion LMWH could ameliorate hypercoagulability following severe burns.

Objective To investigate the influence of composite transplantation of two kinds of acellular xeno-dermis (pigskin) and micro-autograft on wound healing. Methods Seventy-two Sprague-Dawley rats with full-thickness skin defect on the back were randomly divided into three groups, and then compositt skin I(acellular dermal xeno-matrix+autologous particulate skin +selective acellular xeno-dermis) was adopted to cover the skin defect in the experimental group I, composite skin II(acellular dermal xeno-matrix+autologous particulate skin + glutaraldehyde xeno-skin) was adopted to cover the skin defect in the experimental group II, and auto-particulate skin was adopted in the control group. The area of wound healing and the rates of wound contraction were calculated.Tissue samples were harvested and examined by means of histology, immunohistochemistry and electron microscopy. Results There were continuous epidermis and basement membrane, and the dermal matrix was a kind of matrix with normal structure and organization of collagen and without any cellular components in selective acellular xeno-dermis. There was full differentiation of epithelium, orderly collagen arrange and basal membrane of the skin could be identified by immunohistochemical stain in compound skin grafting group I. The healing effect of compound skin grafting group I was better than compound skin grafting group II and auto-particulate skin grafting group. Conclusion The composite grafts constructed by selective acellular xeno-dermis (pigskin) could be a potential new type of composite skin substitute for the repair of full-thickness skin defect.

Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic activity of alkaline phosphatase.We anticipate that the ZZ-AP fusion protein has a potential application in immunoassay field.