ABSTRACT
To clone and express 27 kDa cysteine protease (CP) gene of Spirometra erinacei plerocercoid,and analyze the biology characteristics,a total of RNA was extracted from the plerocercoids and reversely transcribed into cDNA.The 27kDaCP gene was amplified by PCR and cloned into pM-19T vector for sequencing.The accurate sequence was subcloned into the expression vector pET-28a (+).The recombinant plasmid was transformed into Transetta (DE3) and the expression protein induced by IPTG.The recombinant protein was purified by Ni2 + affinity chromatography,and analyzed by SDS-PAGE and Western blotting.The 27 kDa CP gene and its expression protein were predicted and analyzed by bioinformatics analysis tools such as NCBI and ExPASy.Results showed that the ORF length of 27 kDa CP gene sequence was 1 011 bp,and the removed signal peptide sequence was 954 bp with the submission number of ANA52569 in GenBank.The whole sequence of 27 kDa CP (Mr 35 669.9,pI 5.92) was 317 amino acids conferred from cDNA,which belongs to the Peptidase_C39_like superfamily.The pET-28a (+)-27kDa-CP was expressed under the induction of IPTG.Western blotting analysis showed that the purified protein reach expectancy,and had better response with positive serum of Spirometra erinacei plerocercoid infection.In conclusion,the 27 kDa CP gene of Spirometra erinacei plerocercoid is successfully cloned and expressed and knowing coded sequences and bioinformatic.
ABSTRACT
Objective To observe the immunoglobulin A secreting cells (IgASCs) and secretory IgA (sIgA) level in jejunum of mice infected with plerocercoids,and to understand the roles of resistance to invasion processes of the plerocercoids.Methods A total of 100 Kunming mice (half males and half females) were chosen,the weight was 20-25 g,they were randomly divided into control and experiment groups according to their body weight via the random number table method,50 per group.Mice of experiment group were fed each with 5 plerocercoids in snakes,and mice of control group were not infected,testing time and methods were the same in the two groups.Ten mice were randomly sacrificed from one group on days 1,7,14,28 and 56 after infection,to collect empty intestinal juice and jejunal segment.The immunohistochemical method was used to examine the quantity of IgASCs in jejunal mucosa,while enzyme-linked immunosorbent assay (ELISA) was applied to test the level of slgA of jejunal fluid.Results The IgASCs were distributed in lamina propria of the jejunal mucosa,and the percentage of positive IgASCs reached the peak value [(64.24 ± 0.60)％] at d 1 after infection in experiment group,then decreased,and it was lower than control group at d 14 [(41.98 ± 0.42)％ vs (43.52 ± 0.94)％,t =-4.727,P ＜ 0.01].The sIgA level reached the peak value [(22.05 ± 1.43) mg/L] at d 7 after infection in experiment group,then decreased,and there was no statistical significant difference between control and experiment groups at d 56 [(20.00 ± 0.42) mg/L vs (21.26 ± 2.59) mg/L,t =1.516,P ＞ 0.05].There was a positive correlation between the percentage of positive IgASCs in the jejunal mucosa and sIgA level in the jejunal fluid at d 7 after infection (r =0.663,P ＜ 0.01),and there was a negative correlation between them at d 14 after infection (r =-0.542,P ＜ 0.05).Conclusion The plerocercoids infection might induce high level expressions of IgASCs and sIgA,they show positive correlation at d 7 after infection.
ABSTRACT
Objective To explore the influence of different injection techniques on the quality of bolus in 99mTc-DTPA renal dynamic imaging.Methods 395 patients accepted 99mTc-DTPA renal dynamic imaging were retrospectively analyzed.All patients were divided into three groups according to injection techniques:direct injection group (187 cases),intravenous route injection group (84 cases)and venous indwelling needle injection group (124 cases).The three groups were injected by each technique.Areas of interest (ROI) were drawn on abdominal aorta by Xeleris workstation in blood flow perfusion imaging.The time-radioactivity curves of ROI were got.The patients whose ROI curve formed a peak was successfully injected,and did not formed was unsuccessfully injected.The number of patients in three groups who were successfully or unsuccessfully injected was respectively calculated.The data of three groups was taken Chisquare test by SPSS17.0 software.Results 174 patients of the direct injection group,46 of the intravenous route injection group and 115 of the venous indwelling needle injection group were injected successfully.The successful rate respectively was 93.0％,54.8％ and 92.7％.The successful rate of the direct injection group and venous indwelling needle injection group were higher than intravenous route injection group.The difference had statistical significance.The successful rate of the direct injection group and venous indwelling needle injection group hadn't statistical significance.Conclusions The successful rates of the direct injection group and venous indwelling needle injection group were similar.The venous indwelling needle injection technique can be chosen.The successful rate of the intravenous route injection group was lower than the other two groups.The intravenous route injection technique should be chosen prudently.
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Objective To gain experimental data about breeding Musca domestica larvae by pig manure and to evaluate the effects and cost-benefit of feeding hens with larvae instead of fish powder. Methods The concentration of fatty acids and trace elements in the larvae bred with man-made fodder or pig manure were measured. The quality of eggs produced by hens fed with the larva powder or fresh larvae was investigated. Results The contents of trace elements (Ca,Fe,Cu,Zn,Se) of the larvae bred with pig manure were higher than those with man-made fodder(P0.05). Conclusion Breeding Musca domestica larvae with pig manure could produce good larva protein and reduce the expenses. The quality of the egg produced by the hens fed with larvae is as good as the quality of common eggs.