Objective:To study the promotion effect of recombinant human acidic fibroblast growth factor ( rh-aFGF) on wound healing in children with surgery or accident injury. Methods: Totally 206 cases of injured children were randomly divided into two groups, the treatment group was given rh-aFGF, and the control group was treated with basic fibroblast growth factor (bFGF). The ob-servation of wound healing, infection and fat liquefaction in the children was recorded in 7 days,and the modification of scar, pigmenta-tion and adverse reactions were observed in 6 months after the treatment. Results:After the 7-day treatment by rh-aFGF and bFGF, the healing rate respectively was 88. 4% and 74. 2%, and the residual wound area was (2. 2 ± 1. 1) cm2 and (3. 9 ± 2. 4) cm2,respec-tively. Six months after the treatment, the scar incidence rate of the treatment group and the control group was 27. 2% and 67. 5%, and the incidence of pigmentation was 30. 1% and 69. 9%, respectively. Conclusion:Rh-aFGF is more effective than bFGF in pro-moting wound healing in children with surgery or accident injury. It can effectively promote wound healing, shorten the healing time, and diminish the area of scar and pigmentation.

Objective To construct cDNA library of the treated Changliver cell by switching mechanism at 5′ end of RNA transcript (SMART) technique and analyze its quality.Methods cDNA of Changliver cell was aquired with reverse transcription polymerase chain reaction (RT-PCR) and long-distance PCR (LD-PCR),then the cDNA library was constructed with SMART cDNA library construction kit.Results Through testing,the high quality cDNA library containing full length cDNA of Changliver cell had been constructed.The titer of the amplified cDNA library was 4.5 × 1010 pfu*ml-1 and the average exogenous inserts of the recombinants was 1.5 kb.Conclusion These results suggest that the Changliver cell cDNA library has a high quality and lays a solid foundation for researching on Changliver cell and screening

Objective To treat femur fracture in neonates using a new homemade harness designed and observe the effec of this harness. Methods Designed a new harness used in femur fracture in neonates,and used this harness in 7(8 femur fracture)csses,including 6 boys(7 femur fracture)and 1 girl,the age from 1 day to 12 days(average 4.7 days).Those cases included proximal thirds(2 cases)and middle thirds(6 cases)of femur fracture.The angle of fracture was from 44°to 83°(average 62.4°)before treatment. Results The angle of fracture was from 0°to 22°(average 14.0°)after treatment using the harness.Hespitalization was from 2 to 3 days.There were no skin sloughing or harness breaking off.All cases were followed up 6 to 36 months(average 21.3 months).All femur fracture healed in good alignment with leg-length discrepancy＜1 cm.Movement of hip and knee was normal. Conclusions The harness is a better method to treat femur fracture of the proximal and middle thirds in neonates.Advantages of it include simply operation,minimal hospitalization,minimal cost,and easy nursing.

Objective To evaluate the changes of rectal morphology and its clinical value in internal rectal prolapse.Methods The rectal morphology of internal rectal prolapse in thirty-one patients with functional constipation and ten normal control subjects was analysed with defecography.The data were analysed with Mann-Whitney Unonparametric test and Fisher's exact test.Results 23 cases and 2 cases with rectal intussusception in patients group and control group respectively were found.Fisher's exact test was P=0.007.In patients group and control group,the thicknesses of anterior intussusception were(13.51 ±9.42)mm and(3.68±2.34)mm,and posterior intussusception were(5.36±3.92)mm and(2.82±0.99)mm respectively;the intussuscipiens diameters were (37.19±11.79)mm and(25.32±9.25)mm,the intussusceptum lumen diameters were(14.91±4.74)mm and(19.73±6.36)mm,the ratio of intussuscipiens diameters and lumen diameters were 2.82±1.64 and 1.28±0.12,the maximum rectal diameter were(48.97±9.55)mm and(39.84±8.45)mm at rest state.There were significant differences between patients group and control group in above varied values(P＜0.05).Conclusion Defecography can differentiate whole thickness rectal intussusception from rectal mucosal prolapse,which provides the scientific basis for choosing the reasonable treatment.However,caution is required when selecting patients for treatment interventions based on defecography.

ObjectiveTo develop an antibody-array system for multiple detection of antibodies against Japanese B encephalitis virus (JEV),Tick-borne encephalitis virus (TBV),Dengue virus ( DENV ),West Nile virus (WNV),Western equine encephalitis virus (WEEV) and East Equine encephalitis virus (EEEV).MethodsRecombined antigens were spotted on array as capture antigens.Specific antibodies were detected by using a sandwich ELISA format.Rabbit antiserum was employed to select and confirm the specificity of antigens and to optimize the conditions of the assay.The detection efficiency of the system was validated by 40 clinical suspected serum samples and compared with the relative ELISA assays.ResultsEleven recombined antigens were selected as diagnostic antigens with high specificity.Better detection could be achieved when scale of antigen concentrations were within 0.125-0.900 mg/ml and the serum dilutions were 1:100-1:1000.When detecting the 26 clinical suspected TBE serum samples,20 were IgG positive (76.9％),and 17 were IgM positive (65.3％) which was 96.1％ and 84.6％ consistent with the relevant ELLSA tests,the 8 clinical suspected JEV serum samples,4 were IgG positive (50.0％),and 5 were IgM positive (62.0％),which was 86.3％ and 90.1％ consistent with the relevant ELLSA tests.As for the 22 DEN serum samples,13 were IgG positive (60％) and 15 were IgM positive (68％) which was 85％ and 93％ consistent with ELISA.The specificity of the assay was 100％ and the sensitivity was higher than the relative ELISAs.ConclusionThe developed antibody-array is highly specific and reliable,which could be used for the detection of antibodies against the 6 arboviruses.

Objective To express and purify the recombinant nucleoprotein fragments of hemor-rhagic fever with renal syndrome ( HFRS) virus and to evaluate their diagnostic efficacy by using array-ELISA technology .Methods The target genes encoding nucleoprotein fragments of HFRS virus were amplified by PCR, and then inserted into prokaryotic expression vectors to construct the recombinant plasmids of pET -32a (+)/Pn and pET-32a(+)/Pc.The plasmids were transformed into E.coli BL21 ( DE3) to induce the ex-pression of nucleoprotein fragments by IPTG and the expressed products were purified by affinity chromatog -raphy using Ni-NTA agarose.The specificity and sensitivity of the recombinant antigens were evaluated by the assay of array-ELISA using commercial colloidal gold assay kit as a comparison .Results The recombi-nant nucleoprotein fragments of HFRS virus were correctly expressed in E.coli and highly purified by affinity chromatography .Array-ELISA showed that 13 of 16 suspected serum samples were positive by using the His-Pn protein as diagnostic antigen , consistency with the commercial colloidal gold assay kit reaching 94%. Conclusion The recombinant His-Pn protein expressed in E.coli cells could be used for specific serodiag-nosis of HFRS virus as its high antigenicity and sensitivity .The array-ELISA is an effective assay for the de-tection of virus at protein level .

Objective To establish a one-step recombinase polymerase amplification (RPA) method for pathogen screening and rapid detection in the field targeting for five hemorrhagic fever related viruses (Zaire ebola virus, Sudan ebola virus, Marburg virus, Lassa virus and Yellow fever virus). Methods The specific nucleic acid (NA) fragments of each virus were selected as target genes by genome sequence analysis, and the primers and probes for RPA assays were designed according to the sequence. A series of diluted template genes were used for RPA detection to determine the sensitivity. The hemorrhagic fever-related viral nucleic acids were used for RPA detection to determine the specificity. The amplification experiments were carried out at different temperature ranging from 37℃ to 42℃ to validate the reaction temperature range. Results The RPA reaction systems of the five hemorrhagic fever viruses could effectively amplify the target genes, the sensitivities were between 1.5×102 and 1.5×103 copies. No cross reactions existed with the other hemorrhagic fever-related viral genes. Meanwhile, RPA assay could effectively amplify the target genes at 37-42℃. Conclusion The isothermal RPA assays of five hemorrhagic fever viruses are established, which may amply target genes fast and react at a wide temperature range, and be potentially useful for in field pathogens detection.

Objective To investigate the neuroprotective effect of sophocarpine against transient focal cerebral ischemia via down-regulation of the acid-sensing ion channel 1(ASICl) in rats.Methods Twenty-five SD rats were randomly allocated into sham operation,cerebral ischemia/reperfusion,and 5,10,and 20 mg/kg sophocarpine pretreatment groups (n=5 in each group).A rat focal ischemia model was induced by the intraluminal suture method.Five,10 and 20 mg/kg sophocarpine were injected intraperitoneally for pretreatrnent.2,3,5-triphenyltetrazolium chloride staining was used to detect cerebral infarct volume.TUNEL staining was used to detect apoptosis.Immunohistochemistry and Western blot were used to detect the expression of ASIC1 and ASIC2.Results The infarct volume after ischemia-reperfusion was(181.21±9.21)mm3,while the 5,10,and 20 mg/kg sophocarpine pretreatment groups were(150.12±6.19),(52.31±4.20),and(32.18±3.82)mm3,respectively;the neurological function scores in the cerebral ischemia/reperfusion group was(3.62±0.36),while the 5,10,and 20 mg/kg sophocarpine pretreatment grows were(3.15±0.36),(1.92±0.18),and(1.85±0.21),respectively;The surviving neurons only accounted for(31.2±2.8)% of the total cell number in the cerebral ischemia-reperfusion group,while they accounted for(51.2±3.7)%,(76.5±2.1)%,and(77.1±4.1)% in the 5,10,and 20 mg/kg sophocarpine pretreatnmat groups.Compared with the cerebral ischemia/reperfusion group,the cerebral infarct volume was decreased significantly in the sophocarpine pretreatrnent groups(all P<0.01),the neurological function scores were decreased significantly(all P<0.01),and the number of apoptotic cells was decreased significantly (all P<0.01).Immunohistochemistry showed that the number of ASIC-1 positive cells in the sham operation,cerebral ischemia-reperfusion,and 5,10,and 20 mg/kg sophocarpine pretreatment groups were(162.5±8.3),(165.1±5.3),(138.3±7.2),(82.1±6.3),and(69.2±5.5)/mm respectively;Western blot showed that the ASIC1 protein expression was decreased sigaificantly in the 10 and 20 mg/ky sophocarpine pretreatment groups (P<0.01),while there WaS no significant difference in the ASIC2 protein expression.Condusions Sophocarpine may play a neuroprotective role for cerebral ischemia-reperfusion injury in rats via down-regulating the expression of ASIC1 protein.

Theexcretionofurinaryandplasmametabolitesofsalbutamolwasstudiedusingultrahigh performance liquid chromatography electrospray time-of-flight mass spectrometry, after a single intragastric gavaged dose administration with salbutamol. The software of Agilent MassHunter Metabolite ID was employed to find and identify the chemical structure of metabolites of salbutamol. Five metabolites from salbutamol were identified. Metabolites identified in swine urine included glucuronide conjugate salbutamol, N-oxide-salbutamol, hydroxyl-salbutamol, methoxyl-salbutamol and dehydrated hydroxyl-salbutamol. Whereas, only the parent drug, glucuronide conjugate salbutamol and dehydrated hydroxyl-salbutamol were observed in swine plasma.

Objective To express and purify Japanese encephalitis virus ( JEV) EDⅢprotein and evaluate the possibility of using it as a candidate antigen in JEV diagnostic kit .Methods PCR primers spe-cific for the gene encoding JEV EDⅢprotein were designed and used to amplify the gene fragment by RT-PCR.The cloned gene fragment was then inserted into pET-30a (+) to construct the recombinant expression plasmid.The transformed E.coli BL21 carrying expression plasmid were induced by IPTG to express JEV EDⅢ protein.The expressed JEV EDⅢprotein and a control antigen of tick-borne encephalitis virus protein were deposited in small spots to set up ELISA microarray .The serum samples from patients with Japanese encephalitis and healthy people were detected by Array-ELISA.The results obtained by Array-ELISA were compared with those by using indirect immunofluorescence assay .Results The gene fragment encoding JEV EDⅢprotein was successfully cloned and expressed in E.coli BL21.The recombinant protein could be used in Array-ELISA assay for the detection of serum samples from patients with Japanese encephalitis and healthy subjects .The results were consistent with those by using indirect immunofluorescence assay.Conclusion The recombinant JEV EDⅢprotein can be used as a candidate antigen for the diagnosis of JEV infection .