Objective To evaluate the therapeutic efficacy of percutaneous transhepatic portal vein catheterization and thrombolysis on superior mesenteric vein thrombosis.Methods The treatment and therapeutic efficacy of 15 cases of patients with superior mesenteric vein thrombosis underwent percutaneous transhepatic portal vein catheterization and thrombolysis from January 2000 to April 2011 were retrospectively analyzed.Results Percutaneous transhepatic portal vein catheterization was performed successfully in 15patients,without pneumothorax,bile leakage and intra-abdominal hemorrhage after catheterization.Eleven patients had good thrombolytic effect,with majority or complete recanalization on superior mesenteric vein,portal vein and splenic vein.The rate of recanalization Was 73.3％,total mortality was 13.3％.The total amount of urokinase was not more than 500 million U,and there was no cases with systemic bleeding.From 6 months to 36months follow-up,there was no increased portal vein system thrombosis and recurrent cases.Conclnsion Thrombolysis technique of percutaneous transhepatic portal vein catheterization is easy to master,and with good effect of local infusion thrombolytic therapy and lower complication rate.It's a selectable treatment for superior mesenteric vein thrombosis.

Objective To study a new and rapid method for detection of anti-histoplasma antibody by dot immunogold filtration assay (DIGFA). Methods DIGFA was developed by coating purified protein derivative of histoplasmin (P-HTPM) on nitrocellulose membrane as membrane antigen and labeling SPA with colloidal gold. Anti-histoplasma antibodies in sera from mice immunized with Histoplasma were detected by DIGFA. Results All immunizedsera with Histoplasma were positive by DIGFA as well as the results detected by ELISA. The sera immunized respectively with Monilia and Penicillium marneffei were negtive by DIGFA while 1 of 8 immunizedsera with Blastomyces dermatitidis and 2 of 9 immunizedsera with Paracoccidioides brasiliensis were found to be positive. Conclusion DIGFA was a valuable and rapid method to detect histoplasmaantibody in serum.

ObjectiveTo explore the safety and therapeutic effect of 8 patients with esophageal Hiatal Hernia treated by laparoscopic hernia repair.MethodsA retrospective analysis was performed on the clinical data of 8 patients with esophageal Hiatal Hernia form Jun.2009 to Jun. 2010.Among the participants,3 conducted 360-degree fundoplication,5 conducted partial(270-degree) fundoplication.Silk sutures were used for the repair of esophageal perforation in 4 patients,and patch repair was used for the other 4 cases.ResultsEight patients were treated by laparoscopic hernia repair,and all of them were cured without postoperative complications.The mean duration of surgery was ( 120 ± 30) min,with average blood loss ( 50 ± 12 ) ml.Patients had a mean postoperative hospital stay of(4.5 ± 2.5 )days.All the patients were followed up for 1 to 2 years,and no case was found to be relapsed.ConclusionTotal laparoscopic hernia repair is minimally invasive,with short recovery course,less pain after surgery,little complication and short hospitalized time.Laparoscopic Hernia repair should be the preferred effective operation method for patients with esophageal Hiatal Hernia.

Objective To investigate the inhibitory effect of baicalein on mycoplasma pneumonia and its the protective mecha -nism in body , and to provide scientific experimental basis for prevention and treatment of mycoplasma pneumoniae infection .Methods The mycoplasma pneumoniae and baicalein treated BALB /c mice lung tissues were stained with hematoxylin eosin (HE), and his-topathological grading .Minimum inhibitory concentration ( MIC) of baicalein on mycoplasma pneumonia was determined by real-time quantitative polymerase chain reaction ( PCR) .The expression of P 1 adhesion molecules mRNA and protein in lung tissue of BALB /c mice was determined with reverse transcription-PCR and Western blot .The expression of epidermal growth factor ( EGF) mRNA and protein in lung tissue was detected by quantitative RT-PCR and immunofluorescence .Results Baicalein significantly reduced the my-coplasma treated mice's lung tissue pathological score .The minimal inhibitory concentration of baicalein was 32 μg/ml.Baicalein sig-nificantly downregulated P 1 gene transcription and protein translation , and upregulated EGF gene transcription and protein expression . Conclusions Baicalein shows significant resistance to mycoplasma pneumoniae , and can protect the body against mycoplasma damage by inhibiting the expression of P 1 protein and promoting the expression of EGF protein .

Objective To establish a rat model of superior mesenteric vein thrombosis by vein ligation and to simulate the pathological process of the disease, and to provide the basis for studies of its pathogenesis and treatment.Methods Ninety-six SPF male SD rats were randomly divided into three groups: Group A (sham operation group), group B (strangulation group) and group C (simple group), 32 rats in each group.Rats in group A were only opened the abdominal cavity but not blocked the blood supply.The rats were sacrificed at 8, 24, 48 and 72 h after operation.The rats in groups B and C were subjected to establish the strangulation and simple models by superior mesenteric vein thrombosis, respectively, and were sacrificed at 8, 24, 48 and 72 h after modeling.Histological changes (H&E staining) in the rat intestinal tissues were evaluated by a pathological scoring system.The levels of intestinal fatty acid binding protein (IFABP) and α-glutathione S-transferase (α-GST) were detected by ELISA.Results The rat model of mesenteric vein thrombosis was successfully established, with a success rate of 100% (96/96).The pathological analysis revealed that compared with the group A, different degrees of blood stasis and injuries were observed in the intestinal tissues of groups B and C, and the injury were gradually increased in the group B, while gradually reduced in the group C.The degrees of blood stasis and injury were positively correlated with the scope of ligation.The result of ELISA showed that the serum levels of IFABP and α-GST of the rats in groups B and C were significantly higher than those in group A (P < 0.05), and the degree of elevation was positively correlated with the scope of ligation.Conclusions In this study, the rat model of superior mesenteric vein thrombosis is successfully established by vein ligation.This model is simple and easy to operate with a high success rate, and can be used in related research.

Objective To evaluate three different Chlamydophila pneumoniae recombinant antigens for use in Chlamydophila pneumoniae serodiagnosis. Methods The recombinant plasmids pGEX6p-2/ Cpn0146,Cpn0147 and Cpn0308 were constructed and expressed as GST fusion proteins. The immunogenicity and the immunocompetence of these recombinant protein were analyzed by Western-blot and indirect ELISA. A total of 183 sera samples of patients with respiratory tract infection and 32 sera samples of patients with Chlamydia trachomatis infection were detected with indirect ELISA coated microwell plates with the purified recombinant proteins comparing with SeroCP-TM IgG ELISA kits. The positive recognition rate, sensitivity and specificity of each method were analyzed. Results GST-Cpn0146, Cpn0147 and Cpn0308 were obtained after expression and purification. The titers of the specific IgG antibodies against Cpn0146, Cpn0147 and Cpn0308 were higher than 1:6 400, 1:128 00 and 1:128 00, respectively. When the indirect ELISA was developed to detect the IgG antibody against Chlamydophila pneumoniae in 183 samples, the concordance rate between the indirect ELISA test and SeroCP-TM IgG ELISA kits were 92. 3% (Cpn0146) , 94.5% (Cpn0147) and 96.7% (Cpn0308), respectively. The recombinant Cpn0146, Cpn0147 and Cpn0308 were recognized by 71 (38.8% positive recognition rate), 75 (40.9%), and 82 (44.8%) samples, respectively. The recombinant antigen-based detection assays displayed > 97% of detection specificity and>87%of sensitivity.Condusion GST-Cpn0308 shows a better sensitivity and specificity,which suggests it could be used for developing serodiagnosis kits of Chlamydophila pneumoniae infection.

Objective To establish guinea pig model of type Ⅰ anaphylaxis, isolate the anaphy-lactic antibody(IgE and IgG) preliminarily from sera of sensitized guinea pigs, and investigate its functional characteristics. Methods Animal model was established by sensitizing guinea pigs with OVA and Al(OH)_3, level of antibody was determined by ELISA, IgE and IgG in sera were preliminarily isolated through saturated ammonium sulphate precipitate and affinity chromatograph of Protein A. A continuous passive cutaneous ana-phylaxis test (PCA) was performed by sensitizing guinea pigs individually with IgE and IgG and then challenging at different time , and the variation of blue spots in skin were observed after challenge . Results Concentration of IgE in model group and control group were 719.3750 ng/ml and 2.5250 ng/ml, the optical density of IgG in model group and control group were 0.9921 and 0.0174, the level of two antibodies in model group were significantly higher than that of control group (P<0.05). In 9 d continuous PCA test, the blue spot induced by IgE in skin lasted for 9 days and appeared the largest when challenged at day 5. The diameter of blue spot induced by IgG was the largest when challenged at day 2 and then decreased fast. Con-clusion Anaphylactic antibodies were successfully preliminarily isolated from sera of sensitizing guinea pig, both IgE and IgG play roles in type Ⅰ anaphylaxis of guinea pig, the hypersensitive reaction induced by IgG is fast and short than that induced by IgE, and IgG may become an important surrogate marker in immunotoxic-ity evaluation(type Ⅰ anaphylaxis)of vaccine.

ABSTRACT:The guinea pigs were immunized by the Brucella vaccine through intradermal injections and the skin scratch respectively ,and then the immune effects of the two ways were evaluated .Serum samples were collected one month after the last injection and detected for the total IgG titer by interval ELISA .Cell‐mediated immune was evaluated by late‐onset hyper‐sensitivity .The guinea pigs were challenged with Brucella melitensis M5 ,and then were killed to isolated M5 from spleen of each guinea pig to compare the protective effects of two methods of immunization .The ELISA results showed that both of the two methods of immunization could induce strong humoral immune response ,and DTH response to Br‐PPD antigen were 100%in both methods .No significant difference in the immune protective effect of two methods was detected .Results of humoral im‐munity ,cellular immunity and protective effect showed the same effect by intradermal injections and skin scratches .

Objective To comparatively evaluate the effects of a recombinant Mtb protein ESAT 6-CFP10 ( rESAT6-CFP10 ) and a purified protein derivative ( PPD ) as skin test reagents in guinea pigs . Methods Guinea pigs were sensitized with different Mycobacteria species .After sensitization , all guinea pigs were intradermally injected with rESAT6-CFP10 and PPD.At 48 h after the injection, the size of ery-thema at injection sites was measured by using a double-blind method .For guinea pigs sensitized with viable Mtb, the size of erythema at injection sites were measured at 24 h after the injection .The positive conversion rates of skin test with rESAT 6-CFP10 and PPD were calculated .Results The results of PPD skin test were positive in all guinea pigs sensitized with viable Mtb , killed Mtb and BCG with erythema diameters of (11.4 ±0.9) mm, (11.8±1.1) mm and (13.2±0.8) mm, respectively.Positive skin test with rESAT6-CFP10 was only observed in guinea pigs infected by viable Mtb-showing erythema diameters of (13.7±5.7) mm. The skin test with rESAT6-CFP10 was negative in guinea pigs sensitized by killed Mtb-and vaccinated by BCG.The skin tests by using rESAT6-CFP10 and PPD were performed on randomly selected guinea pigs at ninth day after infection by Mycobacterium tuberculosis H37Rv.At the 2nd week, totally 24 selected guinea pigs showed positive skin test results with rESAT6-CFP10 (24/24) with erythema diameters of (19.9± 3.0) mm, while only 15 out of 24 had positive PPD skin test with erythema diameters of (6.1±5.5) mm. At the 4th week, all guinea pigs showed positive PPD skin test (3/3) with erythema diameters of (12.7± 2.5) mm.Conclusion The skin test by using recombinant ESAT 6-CFP10 protein can effectively distin-guish viable Mtb infection from BCG vaccination and killed Mtb sensitization , which is a more suitable anti-gen than PPD for the early diagnosis of Mtb infection .

Objective To establish a suitable guinea pig model for evaluating the protective effects of new therapeutic TB vaccines in preclinical study .Methods The guinea pigs were subcutaneously injected with single-cell suspension of multi-drug resistant M.tuberculosis at a dose of 1000 CFU, 0.2 ml per animal.The study was divided into experimentⅠand experimentⅡ.In experimentⅠ, the guinea pigs were given immuno-therapy and/or chemical treatment on day 3 after infection .Four guinea pigs in each group were dissected at weeks 5, 7 and 9 after infection for evaluating lesion scores and histopathology changes of liver , spleen and lung, as well as bacterial load in spleen .In experimentⅡ, the guinea pigs were given immunotherapy and/or chemical treatment with different doses on day 14 after infection .All guinea pigs were dissected at week 5 after infection for evaluating lesion scores of liver , spleen and lung , as well as bacterial load in spleen .Results In experimentⅠ, all of the three treatments including immunotherapy combined with chemotherapy , immunotherapy alone and chemotherapy alone could effectively prevent organ lesion and reduce bacterial load in spleen , which were significantly different from negative control group .The immunotherapy combined with chemotherapy showed better treatment effects than other two treatments .Along with a prolonged period after drug withdrawal , the de-gree of organ lesion in immunotherapy group and chemotherapy group rebounded sharply , but only slightly in im-munotherapy combined with chemotherapy group .In experimentⅡ, animals in all treatment groups showed alle -viated organ lesion and reduced bacterial load in spleen .A relatively better treatment effect was observed in im-munotherapy combined with chemotherapy group .Conclusion The established guinea pig model of M.tubercu-losis infection showed an advantage of good repeatability .It might be used to evaluate the protective effects of new therapeutic vaccines against tuberculosis in preclinical study .