Objective To investigate the metabolic rules of surfactant protein A and D(SP- A,SP- D )in bronchoalveolar lavage fluid(BALF)in human fetal lungs during gestational ages. Methods BALF with 30 mL saline was performed on clinically collected human fetus with induction of labor by water- bag Their BALF was respectively retrieved [total retrieval rate(85. 6% ? 13 1)% ]for analysis of protein content. The BALF SP - A and SP - D from fetus of various gestational ages or newboms were detected by RPHA and ELISA. Results The total protein in BALF gradually increased since 10th week to newborn peak during lung development. And SP - A and SP D were respectively updated from(0.34 ?0.07 ) ,(0.05?0.01) ng/L to newborn climax[ (6 42 ? 0 36),(1.22 ? 0 13)ng/L] .Conclusions The protein in BALF gradually increases with fetal growth and lung development. SP-A and SP- D may reach prenatal climax and become the main indicator of newborn lung maturity.

Objective To study the way of quick identification of haemophilus influenzae with reverse dot blot.Methods Oligonucleotide probe which is specially targeted to 16SrDNA of haemophilus influenzae was designed, then fixed the probe to nylon membranes, and hybridized with the production of gain with the universal primers.Results The universal primers could hybridize the target sequence from common pathogenic bacteria by PCR, and oligonucleotide probe could hybridize with haemophilus influenza specially and could not hybridize with other bacterias. It proved that the probe was of highly speciality.Conclusion Reverse dot blot is a good method of quickly identification of haemophilus influenzae.

Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

Objective To explore the effect of hyperoxia through mechanical ventilation induced lung injury(VILI) in newborn rabbit.Methods One hundred newborn rabbits aged 1-5 days were randomly divided into 4 groups: with 900 mL/L O2 and mechanical ventilation(MV),group A received high peak inspiratory pressure(HPIP),group B received moderate peak inspiratory pressure(MPIP),group C received low peak inspiratory pressure(LPIP),and group D with no mechanical ventilation with room air.There were 30 rabbits in group A,B,C,10 rabbits in D group.All rabbits were killed at 1,3,6 h after trial respectively.Wet-to-dry weight ratio(W/D) of left lung,and white blood cell(WBC)counts in broncho alveolar lavage fluid(BALF) were measured.The changes of lung histopathology were assessed by hematoxylin-eosin(HE) staining and observed under light microscope.Results Compared with group B and C,the group A demonstrated more pulmonary hemorrhage and pulmonary bullae formation,and the WBC and neutrophils counts in BALF increased and the W/D was higher compared with group B and group C.Meanwhile,alveolar epithelial cellⅡ(AECⅡ) hyperplasy transformed to AECⅠ.In group C,AECⅡswelled and lung tissue edema showed obviously,after 6 hours collagen fibers hyperplasia appeared.Part pulmonary atelectasis was obvious in group C.Conclusions HPIP ventilation can increase lung injury induced by hyperoxia in newborn rabbits,but has minimal effects on MPIP and LPIP ventilation.Pulmonary histopathologic changes participate in the newborn rabbit machine ventilation induced lung injury.

Objective To evaluate the safety and feasibility of minimally invasive percutaneous nephro-lithotomy (mPCNL) with U100plus laser for the treatment of renal calculi. Methods From October 2006 to December 2008 ,mPCNL was performed on 133 patients suffering from renal calculi by using Wolf 8/9. 8 rig-id ureteroscope and U10Oplus laser. Results mPCNL was completed in all the 133 cases. Residual calculi were found in 7 cases after operation and use medical drags to treat. The most residual calculi were removed after 1 month and a few stones being survived. 5 cases with residual calculi were treated by ESWL. The total stone clearance was 91.0%. The operation time was 38 -65 min(mean 46 min). Nephrostomy tube was kept for a mean of 1 -2 d. The mean postoperational hospital stay was 2 -4 d. Among the patients, 133 were followed up for 1-16 months (mean 8. 3 months) , during which no recurrent renal stones were found by B ultrasonngraphy or X ray. Conclusion By using mPCNL with U100plus laser, patients with renal calculi can be treated safely and effectively.

Objective To explore the relationship between the spot size and the result of ploidy analysis in the detection technology based on single-particles, and to fix the scope of spot thickness on the basis of the experimental result.Methods The influence of spot thickness on voltage signals produced by single cells was analyzed.The parameters of the beam shap system in the incident field were designed and optimized on ZEMAX.Finally, according to the cells′diameter, the target size of spots was set.A set of spots of different thickness and of Gaussian distribution obtained from the optical experimental platform was used to conduct ploidy detection experiments.Results Target spots were both obtained from ZEMAX simulation and the optical platform.When the spot thickness was larger than both monocytes and coenocytes, the mean fluorescence intensity ratio was 2.03,which met the demand of the index.Conclusion When the height of the pulse is used to represent the fluorescence density, the relative size of spots and cells will affect the result of ploidy detection.Only when spot thickness is larger than cells is the ploidy ratio accurate.

Objective A major component of flow cytometry data analysis involves gating , which is the process of identifying homogeneous groups of cells .As manual gating is error-prone, non-reproducible, nonstandardized, and time-consuming , we propose a time-efficient and accurate approach to automated analysis of flow cytometry data .Methods Unlike manual analysis that successively gates the data projected onto a two-dimensional filed, this approach, using the K-means clustering results , directly analyzed multidimensional flow cytometry data via a similar subpopulations-merged algorithm.In order to apply the K-means to analysis of flow cytometric data , kernel density estimation for selecting the initial number of clustering and k-d tree for optimizing efficiency were proposed .After K-means clustering , results closest to the true populations could be achieved via a two-segment line regression algorithm .Results The misclassification rate (MR) was 0.0736 and time was 2 s in Experiment One, but was 0.0805 and 1 s respectively in Experiment Two. Conclusion The approach we proposed is capable of a rapid and direct analysis of the multidimensional flow cytometry data with a lower misclassification rate compared to both nonprobabilistic and probabilistic clustering methods .

Objective To investigate the influence of marrow-derived mesenchymal stem cells(MSCs) on intercelluar adhension molecule-1(ICAM-1) in lung of neonatal rats suffered hyperoxia.Methods Marrow-derived MSCs were separated,cultured,amplificated and labeled with 5bromo 2′-deoxy-uridinel(BrdU);after suffered 95% oxygen for 7 days,24 three-day-old SD rats were randomly divided into group A,B and C,and they were injected intraperitoneally with MSCs of 1?10~4,5?10~4 PBS,respectively.Seven days later,immunocytochemisty was used to determine the expression of BrdU and ICAM-1,and value of radical alveolar counts(RAC) of lungs were counted for histopathological study under light microscope.Results Both group A and B,the labeled MSCs had been(detec)-ted in lungs,and there existed significant variance between two groups(P

Objective To investigate the effect of matrix metalloproteinases(MMPs) in ventilator-induced lung injury inflammatory reaction of newborn rabbits.Methods Seventy-five newborn rabbits aged 1-5 days were randomly arranged to 3 groups:control group(n=3),other 72 rabbits designed by 2?2?3 factorial were divided into high concentration oxygen(100%)and low concentration oxygen(45%) groups,and each group was subdivided into 2 groups:High peak inspiratory pressure(PIP)group and low PIP group.All rabbits were done by mechanical ventilation and killed to obtain lung tissue at 1,3,6 hours after trailing respectively.The concentration of MMP-9 and MMP-2 in lung tissue bomogenate were detected by ELISA,at the same time the ratio of wet to dry was detected and pathological section was analyzed.Results 1.The concentration of MMP-2 in high concentration oxygen group,high PIP group and low PIP group increased compared with that in normal group,and there were significant differences.2.The mean concentration of MMP-9 was lower in high concentration oxygen group than that in normal control group,and there was significant difference between 2 groups.The mean concentration of MMP-9 in low concentration oxygen group was higher than that in normal group,and there was no significant difference between 2 groups.There was no significant difference in effect to mean concentration of MMP-9 by different PIP.3.There were positive correlation between MMP-2 and MMP-9,MMP-2 and the ratio of wet to dry in lung.Conclusions Within 6 hours in newborn rabbits by mechanical ventilation,ventilation with high concentration oxygen up-regulate MMP-2,but down-regulated MMP-9.The lungs stretch by mechanical ventilation increase MMP-9,but decrease MMP-2.Mechanical ventilation can affect the synthesis of MMPs,and MMPs plays an important role in ventilator-induced lung injury of newborn rabbits.

Objective To explore the roles of clara cell secretory protein(CCSP)and eosinophil cationic protein(ECP)in the pathoge-nesis of bronchial asthma and to evaluate their diagnostic value in asthmatic children.Methods Induced sputum samples were obtained from 31 asthmatic children during chronic persistent period and clinical remission period.According to global initiative for asthma(GINA),the total of 31 cases accepted systemic treatment by inhaling glucocorticoid.The patients included 18 boys and 13 girls aged from 3.7 to 12.0 years,and their average age was 7.6 years.Sputum CCSP concentrations were measured by enzyme-linked immunosorbent assay(ELISA).And the concentrations of sputum ECP were determined with Pharmacia UniCAP system.Results Asthmatic children had significantly lower CCSP levels in sputum during chronic persistent period compared with clinical remission period(P