Objective To study the influence of probiotics on the gut microflora,digestive enzymes and small intestinal propulsive rate of rats with severe head injury. Methods The rat model of severe head injury was prepared, SD rats were randomly divided into group A (enteml nutrition) , group B (enteral nutrition plus probiotics) , and group C (normal diet) . The intestine mucesa and faeces were collected on the third day,7th day and 14th day after head injury in order to detect gut microflora,digestive enzymes and small intestinal propulsive rate. Results Compared with group C, the number of Lactobacillus and Bacillus bifidus group in group A and group B declined in two weeks after trauma, however, the number of Escherichia coli increased significantly. Bacillus bifidus amounts of group A were significantly lower than that of group B at every time point, the diversity of Escherichia coli was opposite. No difference was seen in content of Lactobacillus 7 days after trauma in group A and B. The level of disaecharidases, Na+- K+- ATPase and small intestinal propulsive rate declined significantly at every time point, compared with group C.Though the contents of digestive enzymes was higher in group B than that of group A, and small intestinal propulsive rate was higher in group A and B than that in group C, but the two groups showed no difference. Conclusions Probiotics can alleviate the in-testine microflora disorder,modulate the activities of digestive enzymes, therefore lessen malabsorption of rats with severe head injury.

A kind of colloidal blue dye(D-l)used as a staining reagent for immunoassay was first selected from the dyes produced in China in this study. The optimun condition for labelling the dye onto sheep antihuman IgG antibody was explored. The dye-labelled antibody could react and stain with relative antigens. The minimal concentration of human IgG protein could be detected with the labelled dye by Dot Double Antibody Sandwich Assay was 20-10ng/ ml. 61 of 62 sera samples of schistosomiasis japonica were positive,the positive rate was 98.4%. Just 1 out of 30 healthy people's sera sambles was positive,the false positive rate was 3.3%. All of 40 Fasciolopsiasis sera samples were negative. The results were similar to that of Dot-ELISA. The activity of antibody labelled with colloidal dye could be maintained for 1 week in room temperature and be presered in lyophilized condition for a longer period. The assay was less expensive,simple to conduct and required no special equipment. It suggested that the Dot Colloidal Dye Immunoassay(DIA)might have potential value for use in schisto-somiasis endemic areas.

Background Flickering light is different from the normal light environment.Animal experiment proved that flickering light can induce myopia.But its mechanism remains unclear.Objective This study was to investigate the expression of c-fos gene in retina of myopic C57BL/6J mice induced by flickering light and monocular form deprivation.Methods Ninety clean C57BL/6J mice aged 28-day-old with the similar refraction in both eyes were randomly assigned to five groups.Fifteen mice in the control group were exposed to continuous white light environment.The white flickering light with the frequency of 10,5,2 Hz were used to irradiate the mice respectively in high frequency flickering group (15 mice),moderate frequency flickering group (15 mice) and low frequency flickering group (15 mice),respectively.The right eyes of other 30 mice were monocularly occluded with a semitransparent hemispherical thin plastic shell to establish the form deprivation models and then were exposed to white light environment.The diopter and ocular axial length were measured by murine-specific eccentric infrared photorefraction and A-scan ultrasonography before experiment and two weeks after the treatments.At the end of experiment,the mice were sacrificed by neck dislocation.Mice eyes were enucleated and retinal samples were prepared for the detect of c-fos protein and its mRNA by immunohistochemistry,Western blot and reverse transcription polymerase chain reaction (RT-PCR),respectively.Results Immunohistochemistry showed that the expressing rate ofc-fos protein in retina was (68.000±10.368)％,(51.000±6.519)％,(46.000±6.519)％,(31.000±7.416)％ and (25.000 ± 7.071)％ in the control group,high frequency flickering group,moderate frequency flickering group,low frequency flickering group and form deprivation group respectively 2 weeks after experiment.The expression rates of c-fos protein in retina in different frequencies of flickering light groups and form deprivation group were significantly lower than that in the control group (t =3.104,4.017,6.490,7.661,all P＜0.05),with the lowest rate in the form deprivation group (P＜0.05).The expression of c-fos detected by Western blot assay exhibited that the relative values of c-fos protein in retina (c-fos/GAPDH) was 0.804±0.050,0.687±0.047,0.667±0.036,0.558±0.036 and 0.532 ±0.056,respectively in the control group,high frequency flickering group,moderate frequency flickering group,low frequency flickering group and form deprivation group,illustrating significantly lowing in different frequencies of flickering light groups and form deprivation group compared with control group (t =2.961,3.184,6.971,6.276,all P＜0.05),whereas the c-fos in the low frequency group and form deprivation group,c-fos protein was less expressed in comparison with the higher frequency flicking group (P＜0.05).The expression level of c-fos mRNA (c-fos mRNA/GAPDH mRNA) in retina was 0.820±0.056,0.663±0.061,0.627±0.034,0.521±0.041 and 0.474 ±0.045 in the control group,high frequency flickering group,moderate frequency flickering group,low frequency flickering group and form deprivation group,respectively.These results demonstrated a significant decline in the expression of c-fos mRNA in different frequencies of flickering group and form deprivation group compared with the control group(t=3.262,5.070,7.173,8.305,all P＜0.05),and the inhibition ability of low frequency of flickering group and form deprivation group was much stronger.Conclusions The c-fos gene level in the retina has a negative relationship with the severity of myopia induced by flickering light and form deprivation.

Objectives To express the miracidial antigen from eggs of Schistosoma japonicum (Chinese mainland strain) (SjMP10), and investigate the role of the miracidial antigen during the hepatic granuloma formation of schistosomiasis. Methods A pair of specific primers was designed and synthesized according to the nucleotide sequence of the open reading frame of the miracidial antigen gene. The open reading frame of the miracidial antigen gene was amplified, digested by restrictive enzyme(BamHI, SalI), and cloned directly into the expression plasmid pGEX-4T-3 to construct the recombinant plasmid. The recombinant plasmids were transformed into E. coli BL21(DE3), and induced by IPTG to express the fusion protein of GST-SjMP10. The expressed fusion protein was purified by electric elution method, and its antigenicity was examined by Western blotting and lymphocyte proliferation test. Results The gene of miracidial antigen was cloned into the expression plasmid pGEX-4T-3. After induced by IPTG, the recombinant expressed a fusion protein of GST-SjMP10, with a molecular weight of 39 000 approximately. The purified fusion protein showed proper antigenicity that could be recognized by the sera of rabbits heavily infected by Schistosoma japonicum and could stimulate the proliferation of splenic lymphocytes of infected BALB/c mice. Conclusion The miracidial antigen from eggs of Schistosoma japonicum was expressed successfully.

OBJECTIVE To analyze the resistance feature and the distribution of Enterococcus species isolated from bile specimens.METHODS Totally 715 samples of bile in intra-operation were cultured,and the drug resistances were tested.RESULTS From them 511 strains in 469 cases were detected out(65.6%),among whith 156 were Enterococcus strains and 159 were strains of Escherichia coli.The mixed infection with two kinds of germs was found in 35 cases.The results of drug susceptibility showed the resistant rates of Enterococcus faecalis to penicillin and high concentration gentamicin were lower than those of E.faecium.None of E.faecalis,and E.faecium resisted to vancomycin.CONCLUSIONS The detective rate of the Enterococcus is high in bile samples.The results reveal a different resistance to common antibiotics,but a high sensistivity to vancomycin and teicoplanin.The result of antibiotic susceptibility testing guides rational use of antibacterial agents.

Objective To investigate the value of recombinant fusion protein (GST-HD)of the large hydrophilic domain (HD) of 23 kDa membrane protein of Schistosoma japonicum with the Glu-tathione-S-transferase (GST) of S. japonicum for early diagnosis of schistosorniasis. Methods The rabbits were infected with cercariae of S. japonicum Chinese mainland strain (300 per one). The rabbits' sera before infection and after being infected at different time were collected. The antibodies IgG(s) against recombinant GST-HD and SEA were measured respectively by ELISA to observe the rabbits' immune reaction status to GST-HD and SEA at different time after being infected with the cercariae. At the same time, 90 serum samples of patients with acute schistosorniasis and 30 samples of healthy persons were checked with GST-HD to evaluate its value for early diagnosis of schistosorniasis. Results At the 17th, 21st and 24th day after infection, the positive rates of antibody IgG of rabbits sera against GST-HD were 42.85%, 92.80% and 100.00% respectively, but the positive rates of antibody IgG against SEA were 14. 28% , 50. 00% and 84.60% respectively. The sensitivity of GST-HD for detecting early schistosome infection was higher than that of SEA significantly. The predictive values of positive and negative of GST-HD for detecting acute schistosorniasis was 98. 89% and 96.67%,respectively, and the diagnostic efficacy was 98. 33%. Conclusion The recombinant GST-HD fusion protein has high early diagnostic value for schistosorniasis.

Objective To screen the bacteria and its components which are toxic to Oncomelania hupensis. Methods The samples of Oncomelania hupensis on the point of death and the soil around the snails were collected. The bacteria existing in the snails and soil were isolated and identified by using regular methods. After being fermented, the toxicity of the bacterium components including the ferment supernatant, split products and bacteria to the snail were tested by the toxicity test. Results Totally, 104 strains of bacteria were isolated from the snails and soil samples, which included Gram positive cocci or bacilli, Gram negative cocci or bacilli. The fermenting supernatant, splitting products and 10 strains of bacteria showed different level of toxicity against the snail respectively. All the fermenting supernatant of bacterium PY1-1, PY7-2, PY3-5, PY19-3, PY2-2-2, PY8-2-2, PY16-1 could kill more than 50 percent of snails. Conclusion The bacteria which are poisonous to snails have been isolated that make a good basis for identifying single toxic component against snails.

Objective To evaluate the molluscicidal effect of colistin E on Oncomelania hupensis. Methods The molluscicidal effect of colistin E on O. hupensis was checked by using the immersion method and spraying method. The toxicity of colistin E on fish was observed by using the toxic test. Results The snail mortality of each group was 100% when the snails were immersed in the colistin E solutions at a concentration of 5. 0, 1. 0 g/L for 24, 48 h and 72 h separately. When the snails were immersed in the colistin E solution at a concentration of 0. 5 g/L for 24, 48 h and 72 h, the death rates were 95% , 100% and 100% respectively; when the snails were immersed in the colistin E solution at a concentration of 0. 1 g/L for 24, 48 h and 72 h, the mortality was 90%, 95% and 100% respectively; when the snails were immersed in the colistin E solution at a concentration of 0. 01 g/L for 24, 48 h and 72 h, the mortality was 70%, 86% and 100% respectively. The snail mortality by the spraying method in a dose of 35, 70, 140 mg/m2 of colistin E was 60%, 100% and 100%. The result of toxic test showed that the toxicity of colistin E on fish was low. Conclusion Colistin E is an effective molluscicide, and is worthy of investigation further.

Objective To analyze the full length cDNA sequence of Schistosoma japonicum egg miracidia genes harboring signal sequence.Methods The gene specific primers were designed and synthesized according to S.japonicum egg miracidia cDNA fragment containing signal sequence identified by signal sequence trapping method previously. The 5′and 3′ end cDNA fragments of each egg miracidia cDNA fragment harboring signal sequence were amplified by nest PCR using the first strand cDNA of S.japonicum as the template. The specific PCR fragments were cloned by TA clone method and sequenced. The full length cDNA sequence of each gene with signal sequence was constructed by comparing the cDNA sequence identified with signal sequence trapping method and the 5′ end sequence, the 3′ end sequence and deleting the overlapping fragments. The splicing model between mRNA of signal sequence and one of mature portions of S.japonicum egg miracidia gene was checked by analyzing the genomic DNA sequence structure of some genes with signal sequence. Results The 5′ and 3′ end cDNA fragments of sixteen among thirty cDNA fragment with signal sequence were amplified successfully, and their DNA sequences were determined. The full length cDNA sequences of sixteen egg miracidia genes were obtained by sequence matching and splicing. The results of deduced amino acid analysis found that the signal peptide of gene SjP4001 was the same to the one of SjP1531 and the signal peptide of gene SjP1183 was similar to the one of gene SjP3742. It confirmed that different genes could share the same or similar signal peptide. The data of S.japonicum genomic DNA sequence analysis showed that the S.japonicum could obtain its signal sequence by alternative splicing model or trans-splicing model. Conclusions The full length cDNA sequences of sixteen S.japonicum egg miracidia genes with signal sequence have been defined, it indicated that the S.japonicum egg miracidia genes could get its signal sequence by alternative splicing model or trans-splicing model was found in this study.

Objective To evaluate the diagnostic value of shear wave elastography on deep venous thrombosis in different stage.Methods Models of rabbits'deep venous thrombosis have been established to noninvasively measure Young's moduli of thrombosis in day 1 ,4,7,10 and 14 respectively by shear wave elastography.Results The elastic modulus of thrombosis increased with increment of the stages of thrombosis.Specifically,the mean Young's moduli of thrombosis increased from (4.0±1 .42)kPa (day 1)to (25.71 ±6.09)kPa (day 14),with statistical differences between each two observation time points (P <0.05).Conclusions Shear wave elastography can effectively reflect the elasticity of thrombosis in different stages,and the stage of thrombosis can be estimated judging by quantitative ultrasound elasticity preliminarily.