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Objective To investigate the expression of Wnt-5a gene in primary hepatocellular carcinoma (HCC) and to expose its role and clinical significance in the development of HCC.Methods Real time quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed in 26 fresh HCC samples and the corresponding para-carcinoma tissues to detect mRNA expression of Wnt-5a gene.Wnt-5a protein was detected with immunohistochemical method in paraffin embedding tissues of 85 cases of HCCs and the corresponding para-carcinoma tissues,and 15 cases of hepatic cirrhosis.Results RT-PCR analysis showed that Wnt-5a mRNA (0.102 127 ±0.158 620) in the HCC tissues was more than that (0.020 106 ±0.022 075) in the para-carcinoma tissues (P＜0.05).The positive expression rate of Wnt-5a protein in HCC,para-carcinoma,and hepatic cirrhosis tissues were 21.2％ (18/85),81.26％ (69/85),and 86.7％ (13/15),respectively.The positive rate of Wnt-5a was significantly lower in the HCC than in the para-carcinoma and hepatic cirrhosis tissues (P ＜ 0.01).The expression of Wnt-5a was significantly associated with lower tumor node metastasis (TNM) stages and small alpha fetoproteins (AFP) content of blood serum (P ＜0.05).Conclusions The high expression of Wnt-5a mRNA was found in the gene transcription of HCC,while Wnt-5a protein was absent or low in HCC.It was suggested that the roles of Wnt-5a was interfered at the protein level rather than the transcriptional level in the HCC.
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Objective To evaluate the influence of Eperythrozoon infection on human and mouse erythrocytes and to explore the pathogenesis of Eperythrozoonosis. Methods The specific gene fragment of Eperythrozoon was detected by polymerase chain reaction (PCR) from the venous blood samples of five patients infected with Eperythrozoon. The complement receptor type I (CD35) expression on erythrocytes of these five patients was determined by flow cytometry. Thereafter, the Eperythrozoons were purified from human samples and injected into mice through the tail veins. Blood smear microscopy, PCR and transmission electron microscopy were used to assure the successful infection. The hematological indicators of human and mice, such as red blood cell (RBC) count,hemoglobin (Hb) content, hematocrit and superoxide dismutase (SOD) were evaluated. All results were analyzed by t test. Results More than 80% of treated mice were confirmed to be infected with Eperythrozoon successfully. A fragment of 801 bp specific gene of Eperythrozoon was detected by PCR in samples from both infected patients and infected mice, which were not detected in samples from healthy control people or control mice. CD35 was highly expressed on the erythrocytes of infected patients, but not expressed on the erythrocytes of infected mice. Both RBC counts and Hb content dramatically decreased in infected patients and infected mice. Hematocrit and the activity of SOD also slightly decreased in infected patients and infected mice. Conclusions Eperythrozoon can spread between human and mice and destroy erythrocyte structure. Eperythrozoon can upregulate CD35 expression in human, but there is no CD35 expression in mice.
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Objective To prepare VP1 protein vaccine of Coxsackievirus A16(CA16) and evalu-ate immunngenicity the subunit vaccines of Coxsackievirus (VP1), and to establish foundation for studying CA16 vaccine. Methods CA16 VP1 was amplified by RT-PCR and cloned into pFastBac HT A plasmid, recombinated with Bacmid DNA by transposition reaction and then transfected Sf9 cell, mixed with adjuvant AI(OH)_3. After immunization BALB/c mice, evaluating immune effectiveness after booster injections 2 weeks. Results The expressed protein was analyzed by SDS-PAGE and Western blot, mice immunized with CA16 (VP1) both induced specific IgG antibody and neutralization antibody. The best immunization antigen was 20 μg, IgG antibody was 1: 1600, neutralization antibody was 1:250, typical Th1/Th2 immune response was determined by lymphocyte proliferation assay and cytokine analysis. Conclusion The CA16 VP1 gene was cloned successfully and expressed in Sf9 insect cells, CA16 VP1 protein vaccine induced both humoral and cellular immune response, to lay solid foundation for further study on CA16 vaccine.
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Anus fine of low rectal cancer surgery, can make the quality of Life of patients significantly im-proved,but postoperative anastomotic leakage is still the main complication, its occurrence can lead to in-creased perioperative mortality, prolonged hospitalization, increased cost, causing a great deal to patients suffering. In this paper, we review the causes and prevention of low rectal cancer after anterior resection of anastomotic leakage.
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Aim To study the effect of Taurochenodeoxycholic Acid (TCDCA) on immunologic cell in vitro and in vivo in mice. Methods Five doses were used , including 0.043, 0.065, 0.1, 0.153, 0.189 g?kg-1 in vivo and 0.01, 0.05, 0.1, 1, 10 mg?L-1 in vitro. Effects of TCDCA on peritoneal macrophage in mouse in vitro,splenic lymphocyte, normal organism, immunologic functional inhibiting model and nerve-endocrine-immunoregulation system were investigated. Results TCDCA had the obvious immunoregulation effect on normal organisms in mouse. 0.1 g?kg-1 TCDCA had the obvious anastated effects on the immunological inhibiting model made up by the cyclosporine A and cyclophosphamide. TCDCA may affect the peritoneal macrophage and splenic lymphocyte in vitro directly, meanwhile two kinds of immunological cells showed the obvious immunoregulation effects on the whole. 0.1 g?kg-1 TCDCA can increase the hydrocortisone contents of the nerve-endocrine-immunoregulation system in mouse most remarkably. Conclusion TCDCA had the adjustment effect on the immunoregulation function in mouse.
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Objective: To evaluate the pharmacodynamic of clematis. Methods: 1) The relaxation of isolated ileum muscle of guinea pig by clematic was tested. The antagonism of clematis on histamine or Ach induced ileum muscle contraction was tested also. 2) By using the model of turning of trunk, the pain releasing effect of clematis was evaluated. 3) The experiments about anti-inflammation action of clematis were carried out in two models. Results: Clematis relaxed the ileum muscle. It also antagonized the contraction of the muscle caused by histamine and Ach. A single intrapeditional administration of clematis to Kun-Ming mice significantly prolonged the latency and reduced the writhe number at the turning of trunk model; clematis dose-dependently inhibited the mouse ear swelling caused by xylol. It also had anti-inflammation effect at the other model. The results demonstrated that clematis could release the pain, suppress the inflammation and smooth muscle contraction.