Objective To prepare the immunotoxin protein (183B 2ScFvPE38) which might be useful in immuno guided therapy for ovarian carcinoma and study the activity of the protein. Methods The methods of ELISA and cytotoxicity were used to study the immunotoxin after induced with IPTG and the activity of the immunotoxin. Results The expressed fusion proteins were detected mostly as inclusion bodies at high level, and soluble immunotoxins were also observed. The results showed liable activity of antibody part and toxic part. Conclusion The recombinant fusion protein 183B 2ScFvPE38 keeps the activity of both components and might be of great use in the future to deal with ovarian carcinoma. [

Objective\ By inhibiting approach of apoptosis of T cells to observe if it could reduce the attacks of tumor cells upon lymphocytes and its possible role in tumor treatment. Methods\ The expressions of Fas/FasL genes in ovarian carcinoma cells were detected by using flow cytometry and RT\|PCR method.The construction of pcDNA3\|antisense Fas,the constructed vector was transfected into Jurkat cells with lipofectin,the change in expression of Fas gene was determined by flow cytometry.By means of Annexin\|V and MTT the effect of apoptosis in transduced Jurkat cells was investigated,and also using MTT cytotoxic test to investigate how 3AO cells kill Jurkat cells. Results\ Fas/FasL were expressed in 6 ovarian carcinomal cells.The expression level of Fas protein in Jurkat cells transduced with the constructed vector was decreased.Apoptosis was reduced in antisense Fas\|transfected Jurkat cells after anti\|Fas treatment. Conclusion\ FasL expression in ovarian carcinoma may be one of the reasons for ovarian carcinoma to escape immunosurveillance and attacking lymphocytes.Blocking Fas expression in lymphocytes can partially inhibit Jurkat cells apoptosis induced by anti\|Fas and reducing the attacks of tumor cells upon lymphocyte.\;

Objective To investigate the value of human epididymis secretory protein 4 ( HE4 ) combined with CA125 assay in differential diagnosis of endometriosis cyst and ovarian malignant tumor.Methods The level of HE4 and CA125 were measured by enzyme-linked immunosorbent assay (ELISA) in the serum specimens of 46 cases in endometriosis cyst group,36 cases in malignant ovarian tumor group,60 cases in benign ovarian diseases and 50 women in healthy women group.Those results were shown with median level.The normal range were 0-150 pmol/L in HE4 and 0-35 kU/L,which either one was more than the threshold value defined as positive index.The sensitivity of assay was evaluated by receiver operating characteristic (ROC) curve,the relation and value of HE4 or CA125 alone and combination assay in diagnosis of endometriosis was analyzed by Mann-Whitney U test and correlation analysis.Results (1) HE4:the median levels of HE4 were 52.4,51.0,50.0 pmol/L in group of endometriosis,normal control and benign ovarian tumor,which didn't show statistical difference.However,HE4 was 507.5 pmol/L inovarian cancer group,which was significantly higher than those of 3 groups (P < 0.05 ).(2 ) CA125:there were significant different in median level of CA125 was observed as 743.0 kU/L in ovarian cancer,84.9 kU/L in endoemtriosis,15.4 kU/L in benign ovarian disease,and 11.5 kU/L in healthy women (P < 0.05).( 3 ) The single assay:when compared with that in endometriosis group,receiver operating characteristic area under the curve( ROC-AUC) were 0.933 in HE4 alone and 0.821 in CA125 alone assay in ovarian cancer group.The specificity was 95% and the sensitivity was 79.6% and 49.0%.(4) The combination assay:when compared with those in endometriosis group,the ROC-AUC was 0.936,the specificity was 95% and the sensitivity was 81.0% in ovarian cancer.Conclusions Measurement of HE4 could be used in differential diagnosis of endometriosis cyst.And the combination of HE4 and CA125 assay could discriminate ovarian endometriosis cysts from ovarian malignant tumors effectively.

Objective To identify the T cell epitopes from ovarian cancer associated anti-idiotypic antibody 6B11in order to explore the molecular basis of 6B11 induced cellular immune responses against ovarian cancer.Methods Potential human leukocyte antigen(HLA)A0201 ligands were predicted by using SYFPEITHI algorithm and tested by the T2 binding assay for screening of HLA-A2 binding peptides from 6B11 complimentary determining region(CDR).Cytotoxic T lymphocytes(CTL)to 6B11 or peptides were generated by 3 rounds of in vitro stimulation with 6B11 or peptide-pulsod dendritic cells(DC),and then tested by 51Cr-release assay to ascertain the CTL epitope of 6B11.Cell proliferation assay was performed by using 6B11 specific CTL as responder cells and peptide-pulsed DC as stimulators to ascertain the helper T lymphocyte(Th)epitope of 6B11.Cytokine assay and interferon-γ ELISPOT assay were performed to verify the CTL and Th epitope of 6B11 further.Results Light chain CDR3 peptide(VL CDR3)of 6B11 induced specific CTL to kill HLA-A2 and target antigen positive ovarian cancer cells,which could be blocked by anti human major histocompatibility complex(MHC)Ⅰ antibody.Heavy chain CDR3 peptide(VH CDR3)of 6B11 stimulated the proliferation of 6B11-primed CTL,which could be blocked mainly by anti human MHC-Ⅱ antibody,and further experiments showed that part of the VH CDR3 peptide-primed CTL killed K562 cells.Peptides in VL CDR3 and VH CDR3 were the CTL and Th epitope mimicking the original antigen of 6B11 respectively.Collaboration of 6B11 CTL and Th epitope,or 6B11 CTL epitope and keyhole limpet hemocyanin(KLH),the former was more powerful in inducing specific cellular immune responses against ovarian cancer.6B11 or corresponding CTL and Th epitope specific CIL secreted high levels of interleukin-2 (1630,1503 ng/L)and interferon-γ(5620,5421 ng/L),while basal level of interleukin-4 was detected (253,274 ng/L).ELISPOT assay confirmed the existence of specific interferon-γ secreting cells in 6811 or epitopes specific CTL(196/1×106 T cell,184/1×106 T cell).Conclusions VL CDR3 and VH CDR3 peptides of 6B11 are the CTL and Th epitopes mimicking original antigen which could duplicate the activity of intact 6B11 to induce the cellular immune responses against ovarian cancer.The results have significant theoretical and practical value in application of anti-idiotypic antibody ag anti tumor vaccine.The acquired CTL and Th epitopes as constituents of future multiple peptides against ovarian cancer could be used in peptide vaccine based ovarian cancer therapy.

Objective To evaluate the value of human epididymis secretory protein 4(HE4)and CAl25 in the diagnosis of ovariall malignancy.Methods HF4 and CA125 in the serum specimens of malignant ovarian tumor group(30 cases),benign ovarian diseases(110 cases;45 benign ovarian tumor,57endometriotic diseases and 8 pelvic inflammation were included) and healthy women group( 137 cases)were assayed double blindly . The levels and the diagnosis efficiency of the HE4 and CA125 were analyzed.Results (1) The median levels of HE4 and CA125 were significantly higher in malignant ovarian tumor group (244 pmoi/L and 601 kU/L respectively) than those of the benign ovarian diseases group( 32 pmol/L and 22 kU/L respectively)and healthy women group (32 pmoi/L and 11 kU/L respectively) (P =0. 000-0. 029). The median levels of CA125 were also higher in endometriotic diseases and pelvic inflammation groups(53 and 41 kU/L respectively) than those of benign ovarian tumor group and healthy women group (12 and 11 kU/L respectively;P = 0. 000-0. 031 ). (2) The positive rate of HE4 was lower than that of CA12s in malignant ovarian tumor group ( P = 0. 036 ). HE4 was negative in benign diseases and healthy women groups. But the positive rates of CA125 were 56. 1% and 5/8 respectively in endometriotic diseases and pelvic inflammation groups and there were significant differences compared with HE4( P =0. 000). (3)The HE4 assay had advantage over the CA125 assay in receiver operating characteristic-area under the curve (ROC-AUC) and sensitivity with a specificity of 100% when ovarian malignancy was compared with controls having benign diseases and healthy women, benign tumor or benign diseases groups respectively. The CA125 assay had advantage over the HE4 assay in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having healthy women group. (4) Combined assay of HE4 and CA125was better than CA125 alone when ovarian malignancy was compared with controls having any group. (5)Combined assay was better than HE4 alone in ROC-AUC and sensitivity with the same specificity when ovarian cancers were compared with controls having benign diseases and healthy women or healthy women groups. And combined assay was lower in the ROC-AUC and the sensitivity with specificity of 100% than HE4 when ovarian cancers were compared with controls having benign tumors or benign diseases groups respectively. (6) The diagnosis efficiency of the HE4 assay at the level 86 pmol/L determined in ROC curve with controls having benign diseases and healthy women group and at the 95% reference level 50 pmol/L of healthy women or 150 pmol/L recommended by the kit respectively was compared. The sensitivity of 50 pmol/L was 73% higher than 150 pmol/L and 86 pmoi/L, while the specificity and positive predictive value were lower ( P = 0. 002, P = 0. 000 ). The specificity, accuracy and positive predictive value of HE4 assay at the set point of 150 pmol/L and 86 pmol/L were 100%, 96% and 96%. The set point of 86 pmol/L had advantage over 150 pmol/L at the sensitivity of diagnosis, 70% and 63% respectively. But the positive predictive value was 95% lower than 150 pmol/L, being 100%. There was no significant difference( P =0. 883, P = 0. 883 ). Conclusions The specificity of HF4 assay is higher than CA125 assay in the diagnosis of ovarian cancer and HE4 combined with CA125 assay can improve the diagnoses. The set point of 150 pmol/L is advantageous for the accurate diagnosis, while the set point of 86 pmol/L is advantageous for the screening of malignant ovarian cancer.

A series of new monobactam sulfonates is continuously synthesized and evaluated for their antimicrobial efficacies against Gram-negative bacteria. Compound 33a (IMBZ18G) is highly effective in vitro and in vivo against clinically intractable multi-drug-resistant (MDR) Gram-negative strains, with a highly druglike nature. The checkerboard assay reveals its significant synergistic effect with β-lactamase inhibitor avibactam, and the MIC values against MDR enterobacteria were reduced up to 4-512 folds. X-ray co-crystal and chemoproteomic assays indicate that the anti-MDR bacteria effect of 33a results from the dual inhibition of the common PBP3 and some class A and C β-lactamases. Accordingly, preclinical studies of 33a alone and 33a‒avibactam combination as potential innovative candidates are actively going on, in the treatment of β-lactamase-producing MDR Gram-negative bacterial infections.