ObjectiveTo investigate the effect of the immunotherapy of CIK cell, LAK cell and PBLS cell mediated by anti-EGFR/anti-CD3 bispecific antibody (BsAb) respectively on the mice borne human gastric cancer and provide experimental evidence for therapeutic strategy in treating gastric cancer. Methods The mAbs of anti-CD3 and anti-EGFR were cross-linked to prepare the BsAb by chemical synthesis. The experimental therapy on the mice borne SGC7901 human gastric cancer was performed,and then the comparisons of the curative activity among the CIK group, LAK group and PBLS group were conducted in vivo. ResultsThe mean tumor reduction rate of the administration of CIK cells directed by anti-EGFR/anti-CD3 BsAb was(64.9±7.7)％ and higher than those of LAK cells or PBLS targeted by anti-EGFR/anti-CD3 BsAb [(43.5±8.2) ％ and (39.7±6.5) ％] (P ＜ 0.05).The mean tumor weight of the administration of CIK cells directed by anti-EGFR/anti-CD3 BsAb was (473.9±37.7) mg at the end of therapy and was lower than those of LAK cells or PBLS targeted by anti-EGFR/anti-CD3 BsAb [(764.6±88.3) mg and (829.1±104.4) mg](P ＜ 0.05). ConclusionThe CIK cell mediated by anti-EGFR/anti-CD3 BsAb could have better curative effect than other effector cells on gastric cancer in vivo.

AIM:To investigate the effects of glucagon-like peptide-1 (GLP-1) on mRNA expression of SOCS-3 and SREBP-1c in the rats with nonalcoholic fatty liver disease.METHODS:Male SD rats were randomly divided into normal control ( NC) group, high fat ( HF) group and HF+liraglutide ( Lira) group.The rats in HF group and HF+Lira group were given high-fat diet for 16 weeks.After 12 weeks of high-fat diet feeding in HF+Lira group, Lira (600μg? kg-1? d-1 ) was intraperitoneally injected for 4 weeks.At the end of the 16th week, the rats were killed.The pathologi-cal changes of the liver were observed under optical microscope.The serum levels of alanine aminotransferase ( ALT) , as-partate aminotransferase ( AST) , triglyceride ( TG) and total cholesterol ( TC) were detected by automatic biochemical an-alyzer.TG contents of liver were measured by GPO-PAP method.The fasting insulin ( FINS) was determined by ELISA, and insulin resistance index was assessed by homeostasis mode assessment ( HOMA-IR) .The mRNA expression of SOCS-3 and SREBP-1c in the liver tissues was detected by RT-qPCR.RESULTS:Compared with NC group, HOMA-IR, TG of liver, and the serum levels of ALT, AST, TG, TC and FINS in HF group were obviously increased (P<0.01).Compared with HF group, HOMA-IR, TG of liver, and the serum levels of ALT, AST, TG, TC and FINS in HF+Lira group were all obviously decreased (P<0.05 or P<0.01).The mRNA expression of SOCS-3 and SREBP-1c in HF group was signifi-cantly higher than that in NC group (P<0.01).The mRNA expression of SOCSV3 and SREBP-1c in HF+Lira group was significantly decreased as compared with HF group (P<0.05).CONCLUSION:Liraglutide may improve the IR and re-
duce TG of liver through decreasing the mRNA expression of SOCS-3 and SREBP-1c, so as to play a therapeutic role in nonalcoholic fatty liver disease.

Objective To investigate the expression and the clinical significance of Lewis y antigen and Mucin 1 (MUC1),as well as to evaluate the correlation between them in epithelial ovarian tumor.Methods The expression of Lewis y antigen and MUC1 in 60 cases of epithelial ovarian malignant tumors,30 cases of borderline ovarian tumors,30 cases of benign ovarian tumors and 20 cases of normal ovarian tissues were detected by immunohistochemical staining.The relationship between Lewis y antigen and MUC1,and their relationship with biology characteristic of ovarian carcinoma were analyzed.An immunofluorescence double labeling methods was performed to detect the correlation between Lewis y antigen and MUC1.Results In malignant epithelial ovarian tumors,the positive rates of Lewis y antigen was 88.33％,which was significantly higher than the positive rates in borderline(60.00％,x2 =9.6405,P ＜0.01) and benign ovarian tumors(33.33％,x2 =28.8095,P ＜0.01) and normal ovarian samples (0,x2 =52.3457,P ＜ 0.01).The positive rates of Lewis y antigen had nothing to do with the clinical pathological parameters of ovarian tumor,but the expression intensity of Lewis yantigen was increased with the development of the malignant degree(P ＜ 0.05).The positive rates of MUC1 in malignant epithelial ovarian tumors was also significantly higher than that in borderline,benign ovarian tumors and normal ovarian samples (86.67％ vs 53.33％,30.00％,25.00％,x2 =12.0321,29.4064,27.8464 ; P ＜0.01).And the expression intensity of MUC1 also increased with the development of clinical stage(P ＜0.01),but had nothing to do with the lymph node metastasis and histological grade(P ＞ 0.05).In ovarian cancer,both Lewis y antigen and MUC1 were highly expressed,and their expression levels were positively correlated (r =0.707,P ＜0.01),and Lewis y antigen colocalized with MUC1.Conclusion Both Lewis y antigen and MUC1 are associated with the occurrence and development of ovarian cancer.Lewis y antigen and MUC1 might be a sigh of biological behavior in ovarian cancers,and this study provides theoretical evidence of ovarian cancer biological treatment.

· Currently, refresher doctors’training is an important form of continuing medical education in our country, effectively promoting and improving the clinical ability and the service level of practitioners in basic -level medical organization. We combine ophthalmic professional features with refresher doctors’training in practice to explore ophthalmic continuing clinical training specifications.Training on the current status and new changes has been analyzed in order to further improve the training quality. The competent administrative departments and clinical teaching hospital should make full use of its advantages to institutionalize, specialize and standardize the training. We suggest linking the continuing clinical training with job promotion;strengthen core clinical ability and being strict with appraisal system so that after the training the trainees can spread standardized guidelines and technology, alleviating the current medical demand and the difficulties in the reform.

Objective To investigate immunophenotypic characteristics of uterine natural killer (uNK)cells and helper T cell 1/helper T cell 2(Th1/Th2)immunity in third trimester decidua in preeclampsia.Methods The proportions of uNK cell subsets,expression of CD_(69)and CD_(94)on uNK cells and Th1/Th2 immunity in decidua were determined in 20 cases of preeclampsia patients and 11 cases of normal term pregnancies by flow cytometric analysis.Results The percentage of CD_(56)~(bright)CD_(16)~-uNK cell subset in preeclampsia patients and the controls was(17.3?11.1)% vs(17.9?16.8)%,that of CD_(56)~(dim) CD_(16)~+uNK cell subset was(16.3?8.7)% vs(16.2?8.8)%;that of CD_(56)~+CD_(69)~+uNK cells was(37.9 ?18.9)% vs(36.8?19.7)%,that of CD_(56)~+CD_(94)~+uNK cells was(34.9?15.2)% vs(32.7?16.2)% and the ratio of CD_(56)~+CD_(69)~+/CD_(56)~+CD_(94)~+was 1.1?0.2,1.2?0.6.No statistical difference was shown in the above values between the preeclampsia patients and controls.The percentage of cytoto xic T cell(Tc)2 cells was significantly lower in the decidua of preeclampsia patients [(3.0?1.0)% vs(4.3?0.9)%,P= 0.001 ],and the ratio of Tc1/Tc2 in preeclampsia patients was significantly higher than that of normal term pregnancies(17.8?3.4 vs 11.8?4.6;P=0.001);the ratio of Th1/Th2 was increased(15.1?2.4 vs 13.2?3.1;P=0.06).Conclusions The immunophenotypie characteristics of uNK cells do not present any significant change in preeclampsia patients.Owing to Tc2 cell decrease,the Th1/Th2 immunity shifts to Th1 type immunity in the decidua,which might contribute to the pathogenesis of preeclampsia.

Objective To explore the effect of Exosomes isolated from the A549 lung cancer cells on the proliferation of these cells and their ho?mologous tumor cells,HCC827,and the role of the PI3K/Akt and SRC signaling pathways in this process. Methods Exosomes were isolated from the supernatant after density gradient centrifugation of A549 cells. The Exosomes morphology was observed by transmission electron microscopy. The expression of the Exosome?specific proteins was analyzed using Western blotting. Cell proliferation was investigated using the MTT assay. Re?sults The A549?derived Exosomes were 30?100 nm in diameter and had a bilayer membrane.Western blotting showed that CD9 was detected in these Exosomes. The isolated Exosomes promoted the proliferation of the A549 and the HCC827 cells in a dose?and time?dependent manner,ac?companied by the activation of Akt and SRC. Conclusion Exosomes isolated from A549 cells promote the proliferation of the secreting cells and the homologous tumor cells in a dose?and time?dependent manner. The mechanism may be related to the activation of Akt and SRC.

Objective To investigate the effect of Exosomes derived from lung cancer cells on the migration of secretory cells and homologous tumor cells and to explore the role of PI3K/Akt and SRC signaling pathways in this process.Methods Exosomes were isolated from the supematant post density gradient centrifugation of A549,lung cancer cells.Morphology of the Exosomes was studied using transmission electron microscopy.Protein expression was analyzed using Western blotting.Cell migration was analyzed by a transwell assay.Results The double-membrane-bound Exosomes appeared as discal-shaped structures,30-100 nm in diameter.Western blotting showed that CD9 was abundant in the Exosomes.The Exosomes promoted the migration of A549 cells and their homologous tumor cells,HCC827 in a dose-dependent manner,accompanied by the activation of Akt and SRC.Conclusion The Exosomes derived from A549 (lung cancer) cells promote the migration of the secreting cells and the homologous tumor cells.The mechanism may be correlated with the activation of Akt and SRC.

Objective To investigate the expression of inregrin?_3 and integrin?_1 in breast cancer and its bio- logical significance.Methods Immunohistochemical assay was used to determine the expression of integrin?_3 and integrin?_1 in the breast cancer(32 cases).Results In normal breast tissue,the positive expression rates of integrin?_3 and integrin?_1 were 0 % and 25 %.In the breast cancer tissue,the positive expression rates of integrin?_3 and inte- grin?_1 were 36 % and 81%.Conclusion The integrin?_3 and integrin?_1 are close associated with the biological sig- nificance of breast cancer.To examine its expression is useful to evaluate the aggressive degree,metastatic potential and prognosis in patients with breast cancer.

Objective To establish a rapid assay with high sensitivity and specificity based on the sequences for group specific gene (GS) and pathogenicity island pag A gene.Methods The PCR primers and probes were designed after the whole sequence was systemically analyzed with bio-informafion tools and blasted with Genebank database.The amplicons were inserted into plasmids so that they could be used as the standard templates to evaluate the sensitivity of the diagnostic system.This assay was based on TaqMan probes and portable Smartcycle PCR machine.Results The detection level was approximately 100 copies per reaction.There was no cross-reaction with other species of Bacillus.This assay could be completed in one hour in laboratory.Conclusion The duplex TaqMan PCR assay could be used to detect Bacillus anthracis rapidly with high sensitivity and specificity.

Objective　To evaluate the role of transfected angiotensinⅡ(Ang Ⅱ) receptor AT1 anti-sense nucleotide (AT1A) in the expression of subtypes of AngⅡ receptor mRNA, synthesis of protein and nucleic acid in cardiomyocytes. Methods　AT1 cDNA sequence (476 bp) was cloned with RT-PCR and reversely inserted into PcDNA3.1 (5.4 kb) to construct an intact plasmid containing AT1A (PAT1A). The plasmid was then transfected into the cultured cardiomyocytes and identified with RT-PCR and Western blot. The synthesis of protein and nucleic acid identified by 3H-Leu and 3H-TdR incorporation, and expressions of AT1 and AT2 mRNA by RT-PCR, were compared between transfected and nontransfected cardiomyocytes after being stimulated with 10-7 mol/L AngⅡ for 24 h. Results　The plasmid PAT1A were successfully constructed. The AT1 mRNA and its protein were expressed significantly less in the transfected cardiomyocytes than in the control (P<0.01). In the transfected cardiomyocytes, AT1 mRNA expression was markedly decreased, but that of AT2 mRNA obviously increased (P<0.01) when compared with the nontransfected cardiomyocytes after stimulation for 24 h with AngⅡ 10-7 mol/L; no significant difference was found in 3H-Leu and 3H-TdR incorporation between them. Conclusion　After the cardiomyocytes was tranfected with AT1A, the expression of AT1 mRNA was markedly suppressed，while AT2 mRNA up-regulated at the same time. Our results indicate that AT1A blocking can not effectively interrupt the Ang Ⅱ-induced synthesis of the protein and nucleic acid in cardiomyocytes.