Objective:To investigate the inhibitiory effect of CD59-siRNA on CD59 gene expression and the growth of xenografted ovarian cancer in vivo,and to explore the role of CD59 in tumor immune escape.Methods:A2780 cells transfected with siRNA plasmid (T group),A2780 cells transfected with blank plasmid (V group) and the wild A2780 cells (C group) were subcutaneously injected into nude mice,respectively.The inhibitiory effect of CD59-siRNA on tumor growth and CD59 expression was evaluated by tumor growth curves and in situ hybridization (ISH) and immunohistochemistry (IHC).Results:The tumor growth curves demonstrated that the growth of cells transfected with siRNA plasmid was significantly inhibited (P<0.05).ISH and IHC showed that the expression of CD59 mRNA and CD59 protein were decreased significantly compared with other groups (P<0.05).Conclusion:Studies of in vivo experiment demonstrate that CD59-siRNA significantly inhibites the expression of CD59,increases the sensitivity of A2780 cells to the complement attack and inhibites the tumor growth.The results may further indicate the role of CD59 in tumor immune escape.

Objective: To amplify two human mutant CD59 eukaryotic expressing systems and investigate mutant CD59 functional activity. Methods: Mammalian expression vector PLATER of mutant CD59 cDNAs was transfected into CHO together with the pcDNA by lipofectamine,which confered resistance to G418(400 ?g/ml). The positive clones were tested by FIH. Activity of both mutants CD59 was determined by BCECF release assay. Results: Mutant CD59 cDNAs subcloned into the mammalian expression vector PLATER and transfected CHO together with the pcDNA,which confered resistance to G418. The positive clones were tested by FIH.Activity of both mutants CD59 before and after glycation was determined by BCECF release assay,both of them could restrict MAC formation ,and glycation could inhibit CD59. Conclusion: A eukaryotic system that expressing mutant CD59 cDNA was successfully set up.It was found that mutant CD59 could restrict MAC formation,and glycation could inhibit mutant CD59. These would be helpful for the furthur study of link mutant CD59 and the vascular proliferative of diabetes.

Objective To explore the optimal immunohistochemical (IHC)diagnostic index for supraclavicular lymph node me-tastasis in lung cancer.Methods In the premise of conforming to the evidence-based medicine,the biopsy of lymph node in 135 pa-tients with supraclavicular lymph node metastasis was performed and 7 antibodies were selected to be detected.135 cases were di-vided into two classes:class A (110 cases)and B (25 cases).The class A included 14 cases of primary lesion inside the lung by the histological analysis,43 cases of primary lesion inside the lung by the cytological detection and 53.cases of primary lesion inside the lung by the radiographic analysis.The class B included 25 cases of lesion outside the lung by the histological analysis.The detecting antibodies included CK7,CK20,EMA,CEA,TTF-1,SPB and vimentin.Results The sensitivity and specificity of single antibody for deducing the primary lesion inside the lung were 90.0% and 56.0% for CK7,98.0% and 40.0% for CK20,90.9% and 4.0%for EMA,80.9% and 36.0% for CEA,62.7% and 100.0% for TTF-1,65.6% and 100.0% for SPB,and 60.9% and 60.0% for vimentin.The quadruple antibodies (CK7,CK20,TTF-1 and SPB)showed the highest sensitivity(85.0%)and highest specificity (100.0%),which made the relatively high detection rate (80.0%)of lung cancer in clinic.Conclusion Analyzing and researching the IMC results is of great significance to guide the surgical pathological practice based on the evidence-based medicine.

Objective To investigate the value of combination detection of CA15-3,CEA,COX-2 on early diagnosis of breast carcinoma.Methods 68 breast cancer patients,61 benign breast disease patients and 68 healthies were selected.Serum levels of COX-2 were detected by ELISA technique,CA15-3 and CEA levels were detected by electroche-miluminescence technique.Results Serum levels of CA15-3,CEA,COX-2 in breast carcinoma group were (34.67±13.20) U/ml,(7.38±3.87) ng/ml,(43.25±10.87) ng/ml.Their levels were higher than those in benign breast disease group and healthy group,the differences were significant (P ＜ 0.01).In combination detection,the sensitivity was 84.9 ％,and accuracy was 91.2 ％,which were higher than single tumor marker used in breast cancer diagnosis.Conclusion The combined detection of CA15-3,CEA,COX-2 could increase the diagnosis rate of breast carcinoma.

Objective:To search for carrier of short active peptide with binding to human CD59 of the surface of HeLa cell.Methods:Ph. D. 12 display peptide library was biopanning with HeLa cell high-expressing CD59. After competitive test, positive phage clones were identified by sandwich ELISA and sequenced.Results:10 phage clones were identified to have higher combination with CD59.Conclusion:The sequence of peptide with binding to CD59 has been obtained.That might lay a basis for design of short peptide of CD59 on tumor sneaking through.

Objective To explore the values of electromyography to examine birth injuries.Methods 34 cases of birth injuries were studied,the electromyography and motor nerve conduction of brachial plexcy were recorded.Then,the location and severity of birth injuries were determined.Results Among 34 cases,according to the location of injuries,there were 10 cases of upper trunk injuries,7 cases of upper and middle trunk injuries,5 cases of middle and lower trunk injuries,5 cases of lower trunk injuries,and 7 cases of whole brachialplexus injuries.On the other hand,according to the severity of injuries,there were one case of complete injuries and 33 cases of incomplete injuries which included 2 cases of severe injuries and 31 cases of slight injuries.Conclusion Electromyography is the reliable method to identify the location and severity of brachial plexus injuries,and this examination is necessary to diagnose birth injuries with great clinical values.

6).The genes were genes that associated with the regulation of cell differentiation and proliferation,adhesion and signal tranduction,apoptosis,transcription and modulation,and DNA damage and repair that.Conclusion:The differential hybridzation analysis of Atlas cDNA expression arrays can be a useful method for analysing the expression profiles of PBMC genes and studing genes differential expression in SLE and IDDM patients.

Aim To investigate the influence and molecular mechanism of C-phycocyanin(CPC) from Spirulina platensis on apoptosis of HeLa cells in vitro.Methods Firstly,the effect of purified CPC on proliferation of HeLa cells in vitro was determined by MTT assay,and then electron microscope was exploited to observe the characteristic apoptotic features of cells treated with CPC.Subsequently,genomic DNA changes of HeLa cells were observed by agarose electrophoresis.Flow cytometric analysis revealed the influence of different concentrations of CPC on cell cycle of HeLa cells.The expressions of apoptosis related genes of CPC treated HeLa cells were determined by immunohistochemistry analysis.In addition,the activities of caspases and the release of cytochrome c from mitochondria into the cytosol were detected.Results Compared with control cells untreated with CPC,a significant decreased in the numbers of HeLa cells in survival treated with CPC and concentration dose effects existed.CPC could induce characteristic apoptotic features including cell shrinkage,membrane blebbing,microvilli loss,chromatin margination and condensation into dense granules or blocks.DNA of HeLa cells treated with CPC showed fragmentation pattern(DNA ladder of oligomers of 180~200 bp) typical for apoptotic cells.HeLa cells treated with different concentrations of CPC demonstrated an increasing percentage of cells in sub-G0/G1 phase.In addition,CPC could promote the expression of pro-apoptotic gene(Fas and ICAM-1);meanwhile,held back the anti-apoptotic gene(Bcl-2) expression,and then facilitated the transduction of tumoral apoptosis signals that resulted in the apoptosis of HeLa cells in vitro.In CPC treated HeLa cells,CPC treatment of HeLa cells also resulted in activation of caspases and release of cytochrome C from mitochondria into the cytosol.Conclusion C-phycocyanin from Spirulina platensis can induce the apoptosis of HeLa cells in vitro.By virtue of the promotion of the apoptosis signals transduction in HeLa,CPC realizes its antitumor activities.

[ ABSTRACT] AIM:To investigate the change of phosphorylation of tau protein and expression of cyclooxygenase 2 ( COX-2) in colon submucosal neurons of enteric nerve system in inflammatory bowel disease ( IBD) rats induced by tri-nitrobenzene sulfonic acid (TNBS).METHODS:Male rats (n=30) were randomly assigned to 3 groups (n=10 each):control group, IBD group and TNBS group.The IBD rats were induced by TNBS+ethanol enema for 14 d.The control and TNBS rats were given an equal volume of saline and TNBS, respectively.The general situation and the histopathologic change of the rat colon were observed.Immunofluorescence was used to check the change of phosphalated tau protein and COX-2 expression in the submucosal neurons of the colon.The expression of COX-2 and phosphorylated tau231 and tau262 in the rat colon submucosal neurons was observed by double immunofluorescence staining.RESULTS:Compared with con-trol group, the number of neurons in the colon of IBD rats decreased obviously and the expression of phospholated tau231 and tau262 was significantly increased.The number of neurons in the colon of TNBS rats showed no significant difference compared with control rats.The rat neurons in control group and TNBS group did not express COX-2.COX-2 expression was observed in the nucleus and cytoplasm of colonic neurons in IBD rats, which showed significantly different from control and TNBS rats.CONCLUSION:The decreased neurons in the enteric nerve system of IBD rats might be associated with the phosphorylation of tau protein and the expression of inducible COX-2.