[Objective]To investigate the morphological characteristic of the scalenus minimus. [Methods]Totally 32(64 sides) embalmed adult cadavers were dissected and studied,the morphology of scalenus minimus and its relationship to brachial plexus was observed.Ten scalenus minimus were stained by HE to study membrane of the muscles.Twenty-seven(54 sides) embalmed adult cadavers were dissected carefully to investigate its nerve and blood supply.[Results]Scalenus minimus was found in 84.4% of cadavers(54/64).Its insertion was mainly composed of tendinous tissue,which was spaned by the lower trunk of brachial plexus.Scalenus minimus supply nerve branches was from ventral rami of the cervical seven root,and vascular supply was from:(1) branches of deep cervical artery,(2) branches of subclavia artery.[Conclusion]Scalenus minimus muscle,an independent but inconstant muscle,is existed in most people and sometimes responsible for compression of brachial plexus.It is suggested that scalenus minimus muscle should be resected carefully as well as scalenus anticus and medius during surgical treatment of thoracic outlet syndrome.

AIM: To explore the expression of CD14 in rat Kupffer cells (KCs). METHODS: In rat KCs induced by LPS or the mediators from KCs induced by LPS,the changes of CD14 expression were measured by RT-PCR and immunohistochemistry.The expressions of TNF? mRNA?IL-6 mRNA or the concentrations of TNF??IL-6 were estimated by in situ hybridization and radioimmunoassay,respectively. RESULTS: LPS increased the expression of CD14 in KCs in a dose-dependent fashion (LPS,1 ?g/L-10 mg/L) and in a time-dependent fashion(0.5 h-24 h,peaked at 3-6 hours). While the expression of CD14 in KCs stimulated by the active mediators from KCs which had been exposed to LPS 1 hour were obviously increased. CONCLUSIONS: There was a close relationship between LPS or the active mediators from KCs induced by LPS and the expressions of CD14. It is implied that the increase in CD14 expression may be induced by LPS and the cytokines produced by KCs,it also reveals that there is a auto-regulated loop in CD14 expression.

Objective To study the effects of liver graft on the immune tolerance of intestinal allograft in auxiliary en-bloc liver-small bowel transplantation in pigs.Methods Seventy outbreed Landrace pigs were divided into 4 groups.Ten auxiliary liver-small bowel allotransplantations were performed in group A,B,C,respectively,and 5 segmental small bowel allotransplantatiom were performed in group D.Pigs were administered with routine and lower dose of cyclosporine and methylprednisolone in group B and C,respectively. No immunosuppressive agent was administered to pigs in group A and D.Results The initial time of acute rejection was obviously prolonged in group A than group D.and the acute rejection was milder in group A than group D(P＜0.05).There was no significant difference upon postoperative survival time,initial time of acute rejection and degree of acute rejection between group B and C(P＞0.05).Conclusions The immune tolerance of intestinal allograft Can be induced by liver graft in auxiliary en-bloc liver-small howel transplantation.

To explore the effect of estrogen on pathogenesis of osteoporosis in women with maintenance hemodialysis. One hundred and twenty women aged 18-45 years had been undergoing maintenance hemodialysis for ≥ 3 months were included. Of them ,60 women without osteoporosis served as control group and the other 60 women with osteoporosis as observation group. Serum concentrations of estradiol, tumour necrosis factor-α (TNF-α), parathyroid hormone (PTH), and calcium were determined, meanwhile bone mineral density (BMD)was measured by quantitative computed tomography. Serum estradiol levels in the observation group were lower while TNF-α level were higher than those in control group (all P<0. 05). PTH and calcium levels were not significantly different (P= 0.567 and P = 0. 588). In the observation group, linear correlation analysis revealed positive correlation (r = 0. 865 ,P<0. 01)between estradioi and BMD,while multiple linear regression analysis showed that serum estradiol and calcium levels were positively correlated with BMD, and the concentrations of TNF-α and PTH were negatively correlated with BMD (F= 140.32 ,P<0.01). Estradiol levels were found to have greater effect on BMD(t=5. 386, P<0. 01). Lowered serum concentration of estradiol in women with maintenance hemedialysis seems to be a major factor related to osteoporosis,it accelerates the pathogenesis of osteoporosis by modulating TNF-α.

Objective To investigate the effect of raloxifene on ERα, ERβ and MT-1a expression in renal tubular epithelial cells contaminated with cadmium chloride,for exploring the protective effect and action mechanism of raloxifene in renal injury induced by cadmium. Methods Renal tubule cells were isolated from kidneys of new born SD rats and purified by Percoll. The cells of the second generation were selected for experiment. The cells were divided into 3 groups:blank control group,cadmium chloride group and raloxifene group. After 4 h,cell viability was detected by MTT,and after 24 h,mRNA and protein expression levels of ERα, ERβ and MT-1a in renal tissues were determined by RT-PCR and immunohistochemistry technology,respectively. Results Compared with the blank control group,morphology of plenty cells were changed in cadmium chloride group,a number of cells were dead,and the survival rate was only 55.3%;at the same time,mRNA and protein expression levels of ERβ and MT-1a significantly increased (P<0.05). Compared with cadmium chloride group,only a small part of cells underwent morphological changes,and the survival rate was 88.6% in raloxifene group,and the mRNA and protein expression levels of ERβand MT-1a were significantly decreased ( P<0.05) . The expression of ERαshowed no significant difference among the three groups ( P>0.05) . Conclusion Raloxifene can bond with ERβcompetively,down-regulate MT-1a and decrease Cd-MT synthesis,so as to reduce cadmium-induced damage of renal tubular epithelial cells.

Objective To synthesize and fusion express the predicted T-cell epitopes of Schistosoma japonicum, and analyze their immunogenicities. Methods The plus and minus oligo-nucleic acid strands of epitopes P7, P17, P18 were synthesized following their DNA sequence, respectively. The Nco I restriction enzyme sites were added to the 5′ end of epitope gene and the Xho I restriction enzyme sites were added to the 3′ end of epitope gene. The plus and minus strand of each epitope gene was annealed to form double strand DNA fragments. Then the double strand DNA fragments encoding epitope peptide were cloned into the site between Nco I and Xho I of plasmid pET32c(+) to construct recombinant plasmid which was transformed into E.coli DH5?. The recombinant plasmid containing P7, P17, P18 genes respectively was identified by PCR, restriction digestion and DNA sequencing, and then transformed into E.coli BL21 (DE3) for expressing the fusion protein. The fusion protein of peptide-thioredoxin(Trx) was expressed by inducing with IPTG and analyzed with SDS-PAGE. The fusion proteins were purified with Ni2+ column affinity chromatography. Meanwhile, the peptides P7, P17, P18 were synthesized artificially following their amino acid se-quence. By using the purified epitope peptide fusion proteins and synthesized epitope peptides, the splenic cells of C57BL/6J mice immunized with ultraviolet-attenuated cercaria of Schistosoma japonicum were stimulated respectively. The stimulation activity of fusion proteins and synthesized peptides were assayed by detecting the incorporation rate of 3 H-thymidine. Results The double strand DNA fragments of epitopes P7, P17, P18 were successfully cloned to form recombinant plasmids, all of which could express a 20 kDa fusion protein. Both the fusion protein and synthesized epitope peptides of P7 and P17 were able to stimulate the lymphocyte cells to proliferation effectively. Conclusion The peptide P7 and peptide P17 are testified as T-cell epitopes of Schistosoma japonicum.

ObjectiveTo evaluate the app li cation value of the Dipstick Dye Immuno assay (DDIA) for screening chemotherapy targets of schistosomiasis in a lower endemic area. Methods[ WT5”BZ]In a lower endemic area of schistosomiasis a random sample of 463 individuals from a natural village were examined using miracidium hatching metho d, Kato Katz's method, DDIA, DGS COPT and ELISA. The positive rates of these a ss ays were compared. ResultsThe positive rate of stool examination was 3.9% in 463 individuals. The positive rate of DDIA was 15 8%. The positive rate in 18 stool positive subjects was 94 4% with Youden In dex 0 81. The positive rate of DGS COPT was 8 9% . The positive rate in 18 stool po sitive subjects was 72 2% with Youden Index 0 66. The positive rate of ELISA w as 18 4%. The positive rate in 18 stool positive subjects was 83 3% with Youden In dex 0 68. ConclusionDDIA was more suitable for application in screening target population in lower endemic areas than other im munoassys.

Objective To express the cytosolic superoxide dismutase(SOD)of Schistosoma japonicum and analyze its antigenicity.MethodsThe DNA sequence of Schistosoma japonicum gene of cytosolic SOD was amplified by reverse transcription polymerase chain reaction(RT-PCR)and subcloned into the expression vector pGEX-4T-3 to construct a recombinant plasmid Sj SOD-pGEX-4T-3.This recombinant plasmid was transformed into component cell of E.coli BL21(DE3).The fusion protein of GST-SOD was expressed by inducing with IPTG and purification by affinity chromatography.The specific antiserum was prepared by immunizing the BALB/c mouse with GST-SOD fusion protein,and the immnuogenicity of GST-SOD was investigated by Western blot analysis.ResultsThe gene of cytosolic SOD was amplified successfully and subcloned into expression vector pGEX-4T-3 forming the recombinant expression plasmid Sj SOD-pGEX-4T-3.The fusion protein GST-SOD was expressed after the recombinant containing Sj SOD-pGEX-4T-3 being induced by IPTG.Immunizing the BALB/c mouse with the fusion protein GST-SOD produced high titer specific antiserum of 1∶12800 and the fusion protein GST-SOD was recognized by sera of severe infection rabbits.ConclusionsThe cytosolic SOD has been expressed successfully and has a preferable immunogenicity.

Objective To explore the protective effect of recombinant superoxide dismutase(SOD)fusion protein against the infection of Schistosoma japonicum Chinese strain.Methods The recombinant SOD fusion protein was expressed and purified with Glutathione sepharose 4B.C57BL/6J mice were immunized with the recombinant SOD fusion protein mixed with Freund adjuvant.Four weeks after the final immunization,the mice of the experiment and control groups were challenged with(45?2)S.japonicum cercariae.All the mice were sacrificed on the forty-fifth day after the challenge to calculate the worm reduction rate and egg reduction rate,and to observe the pathologic changes of liver tissue of the mice.Results The worm reduction rate was 35.63% and the egg reduction rate was 31.17% in the experiment group.The number of granuloma in the live tissue of the experiment group was less than that of the control group,and the mean diameter of single granuloma in the experiment group reduced by 22.32% compared with that of the control group.The IgG subclass levels of IgG1,IgG2a,IgG2b were higher than those of the control group.Conclusion The recombinant SOD fusion protein has a protective effect against Schistosoma japonicum infection.

Objective To establish rSAG1-IgM-ELISA with purified rSAG1 fusion protein for immunodiagnosis of toxoplasmosis. Methods The rSAG1 fusion protein was purified by Ni 2+ column. The ELISA plate was coated with different concentrations of rSAG1, reacted with pooled positive and negtive human sera. Goat anti-human IgM conjugated to horseradish peroxidase was used as the second antibody. The appropriate detecting condition of the rSAG1-IgM-ELISA assay was determined by orthogonal experiment. The reproducibility, sensitivity and specificity of the assay were assessed. Thirty-five IgM-positive and 57 IgM-negative human sera detected by the imported IgM-ELISA kit were detected with the rSAG1-IgM-ELISA. Results The purity of rSAG1 was above 90%. The appropriate detecting condition was that the coated rSAG1 was 2 5 ?g/ml, the human serum was in 1∶100 dilution, and the second antibody was in 1∶4000 dilution. The coefficient of variation (CV) value of IgM-positive and IgM-negative pooled sera were 13 8% and 7 7% respectively. The inhibition rate of the assay was 62 0% The positive correspondence rate and negative correspondence rate were 82 9% (29/35) and 91 2% (52/57) respectively,the total correspondence rate was 88 0%, compared with the imported IgM-ELISA kit. Conclusions The rSAG1-IgM-ELISA has high sensitivity and specificity, and good correspondence rate with the imported IgM-ELISA kit. It indicates that rSAG1-IgM-ELISA has potential value for early diagnosis of toxoplasmosis.