Objective To screen the tumor specific antigens, and explore new methods for diagnosis and therapy of tumor. Methods Hela cells, the human cervical carcinoma cell line, were used to immunize the BALB/C mice, and the positive hybridomas were screened by rosette formation screening system in human carcinoma cell line Colo205, Burkitt lymphoma cell line, Raji, etc. The immunofluorescence staining and flow cytometry analysis were performed to characterize if the prepared monoclonal antibodies (mAb) could reorganize the natural membrane antigen expressed on Hela, Colo205, Raji, PLA801, 7901, Jurkat, 293T, ECV304 and resting or activated human peripheral blood mononuclear cells (PBMC). Furthermore, the membrane protein of Hela, Raji and Sp2/0 cells was segregated by ultracentrifugation and the mAbs were cross-linked with the Sepharose CL-4B. In order to identify the molecules which could be recognized by the prepared mAbs, immunoprecipitation was performed to purify the tumor antigen candidates with the cell membrane protein from Hela, Raji and SP2/0 (as negative control) through Sepharose CL-4B cross-linked with the prepared mAbs. Finally, western blotting was performed to detect the molecular weight of the products from the immunoprecipitation. Results Three mAbs were obtained which could recognize the tumor antigen candidates expressed in several human carcinoma cell lines, such as Hela, Raji and Colo205, and some embryonic cell lines, such as 293T and ECV304, but they could not bind to Jurkat, PLA801, 7901, SP2/0, resting or activated human PBMC. Western blotting results indicated that the three mAbs could recognize a 47kD molecule which expressed in Hela and Raji but not on SP2/0 cell membrane. Conclusion Three mAbs, which can recognize tumor membrane antigen candidates, are obtained, providing useful tools for screening and identify the tumor specific antigens.

Objective To detect the sequence of SLC29A1 gene rs1288 single nucleotide polymorphism (SNP) in advanced pancreatic cancer patients,and to explore its correlation with gemcitabine toxicity and prognosis.Methods Peripheral blood samples were collected and DNA was extracted.The segment containing SLC29A1 gene SNP (rs1288) was amplified by PCR,and then DNA sequencing was conducted to identify SLC29A1 gene SNP (rs1288).According to the sequencing results,the patients were divided into SLC29A1 gene rs1288 T→A mutation type group and wild type group.Clinical data,toxicity of gemcitabine chemotherapy,progression free survival (PFS) and overall survival (OS) between two groups were compared.Results A total of 83 pancreatic cancer patients were enrolled.Sequencing results showed that SLC29 A1 gene 1288 T→A mutation type was present in 52 patients and wide type was observed in 31 patients,so mutation rate was 62.7％.All the patients in both two groups could tolerate the gemcitabine toxicity,and no chemotherapy related death occurred.There were no statistical differences on the gender,age,CA19-9,tumor site,size and TNM stage between the two groups.There were statistically higher iucidences of leukopenia and thrombocytopenia in the SLC29A1 gene rs1288 T →A mutation type group compared to the wild type group (55.8％ vs 32.3％,P＜0.05;40.4％ vs 19.4％,both P＜0.05).Median OS and PFS in mutation type group were shorter than those in wide type group (11 months vs 14 months,P ＜ 0.05;9 months vs 12 months,P ＜ 0.05).Conclusions Advanced pancreatic cancer patients with the SLC29A1 gene rs1288 T→A mutation type had a higher incidence of adverse reaction in gemcitabine chemotherapy and a worse therapeutic effect,and thus detecting the mutation of SLC29A1 gene rs1288 point mutation may serve as a marker for evaluating the toxicity and prognosis of gemcitabine chemotherapy in patients with pancreatic cancer.

Objective To explore the expression and significance of Annexin Ⅱ protein in ovarian adenocar-einoma.Methods The expressions of Annexin Ⅱ protein were detected in 48 cases of ovarian adenocarcinoma and 10 cases of ovarian adenoma by SP immunohistochemistry.Results The positive rates of expression of Annexin Ⅱprotein in ovarian adenoma were 50.00% (5/10), and were 81.25% (39/48) in ovarian adenocareinoma (χ2 =4.414, P <0.05), in well and moderately-differentiated, poorly differentiated ovarian serous adenocarcinoma were 93.33% (14/15) and 78.95% (15/19) respectively (χ2 = 1.383 ,P0.05 ), and on the Ⅰ and Ⅱ stage, Ⅲ and Ⅳ stage of ovarian serous adenocarcinoma were 60.00% (6/10) and 95.83% ( 23/24 ) respectively (χ2=7.226, P <0.05).The positive rate in the group of ovarian serous adenocarcinoma with lymph node metastasis was 90.00% (27/30), while 50.00% (2/4) in the group without lymph node metastasis (χ2=4.502, P<0.05).Conclusion The positive expression of Annexin Ⅱ protein may be correlated with the carcinogenesis, development and lymph node metastasis of ovarian serous adenocarcinoma.

Objective To study the tissue-specificity of ?-carotene-15,15’-monooxygenase(? CMOOX). Method Semi-quantitative RT-PCR and HPLC were used to study the expression of ?CMOOX mRNA and its activity in different tissues of chicken. Results ?CMOOX mRNA was expressed in tissues including heart, liver, spleen, lung, kidney, duodenum, jejunum, ileum, muscle and testis, not expressed in lung. Furthermore, it was expressed in jejunum with the highest level. ?CMOOX activity was found in every tissue, except lung, and highest in duodenum and jejunum. Conclusion The distribution of ?CMOOX in chicken has tissue specificity.

The mechanism of sudden cardiac death caused by variation in SCN5A is still unclear. Recently, the converging evidences suggest that the dysfunction of regulation mediated by transforming growth factor-β1 in cardiac fibration and reconstruction of cardiac iron channel could be main reason of SUNDS caused by variation of SCN5A. The resent progress of the mechanism of transforming growth factor-β1 in sudden cardiac death caused by variation of SCN5A gene is reviewed in this paper, hoping to provide reference for the research and practice of sudden cardiac death in forensic medicine.

Objective To investigate the effects of dextran sulfate on lung ischemia-reperfusion injury after lung transplantation in rats.Method A total of 32 male Wistar rats were subjected to unilateral left lung orthotopic transplantation.They were randomly divided two groups (n =16 each):DXS group [DXS (10 mg/kg) was given prior to the reperfusion],and the control group (the same volume of normal saline was given).After animals were sacrificed,the lung graft was harvested 2 h after reperfusion.Oxygenation indexes,wet/dry ratio (W/D),myeloperoxidase (MPO) activity,malondialdehyde (MDA) and endothelin 1 (ET-1) in the transplanted lung,and tumor necrosis factor a (TNF-α) and interleukin 8 (IL-8) in serum were measured.The lung injury scores were evaluated and complement deposition was observed.Result After 2-h reperfusion,compared to the control group,oxygenation indexes were improved significantly in DXS group (P＜0.05),but there were no significant differences in W/D between two groups.In DXS group,the activity of MPO was significantly reduced,and the contents of MDA and ET-1 in the lung tissue were significantly reduced as compared with the control group.DXS reduced the level of TNF-α and IL-8 markedly in serum (P ＜0.05).There was no significant difference in lung injury score between two groups (4.53 ± 0.46 vs.5.28 ±0.49,P＞0.05).Compared to the control group,DXS reduced the deposition of C3c (0.8 ±0.2vs1.5±0.3) andC6 (1.2±0.4vs.2.4±0.5) (P＜0.05).Conclusion Administration of DXS attenuated ischemia-reperfusion injury after lung transplantation by inhibiting complement deposition,and improved the oxygenation of the transplanted lung.This protection was associated with inhibition of inflammation and oxidation and endothelial cytoprotection.

Matrix_assisted laser desorption ionization_time of flight tandem mass spectrometry ( MALDI_TOF/TOF MS) and electrospray ionization_quadrupole_time of flight mass spectrometry ( ESI_Q_TOF MS) were used to confirm the structure of cyclic lipopeptide daptomycin fastly. First, the relative molecular weight 1916. 7107 of daptomycin was measured by ESI with error 0. 0007. The sample’s doubly charged peak m/z 809. 848 was selected as precursor ion for ESI_MS/MS analysis, and the exocyclic amino acid sequence C9 H19 CO_Trp_Asn_Asp was successfully matched. Second, the experimental conditions of cleaving daptomycin by lithium hydroxide ( LiOH) were optimized and the ring_opened process was monitored by MALDI_TOF/TOF MS. After obtaining ring_opened product with purity of above 95%, the MS/MS measurements by MALDI and ESI were carried out. The b+and y+of ring_opened product were completely matched, which confirmed the amino acid sequence of daptomycin. Finally, ESI_MS/MS conditions of ring_opened product were further optimized to obtain more low mass fragment ions for analyzing the structure of fatty acid chain and the cleavage pattern of fat chain in mass spectrometry was proposed. The method was fast, convenient, accurate and reliable for identifying cyclic lipopeptide compounds.

ObjectiveTo explore the bone anabolic effects after a single local injection of simvastatin into femoral cavities of osteoporotic rats.MethodsThirty-six female SD rats(3 months old,body weight 250-300 g) were ovariectomized(OVX) and low-calcium-diet fed for 3 months,OVX rats were randomized into 3 groups(n=12).Left femurs of group A,B and C were injected with 0,5 and 10 mg simvastatin,respectively.Half of the rats in each group were randomly euthanized separately 1 and 5 months after simvastatin injection.Left femurs were taken out for bone mineral density (BMD) assessment with dual energy X-ray absorptiometry,bone histomorphometic changes were analysized by Micro-CT,and two kinds of biomechanical tests were used to evaluate the osteogenic effects.ResultsOne and five months after injection,BMD in mid-diaphysis significantly increased in simvastatin-injected groups compared to the control group.For Micro-CT analysis,significant increase in total bone volume/total tissue volume,cortical wall thickness,trabecular thickness,trabecular number,and a significant decrease in trabecular spacing were observed in simvastatin-injected groups compared to the control group.For both biomechanics (the three-pointbreaking test of condyles and axial compressive testing of proximal femur),the values were significantly higher in simvastatin-injected groups than the control group.ConclusionLocal simvastatin treatment showed a positive effect on improving mechanical strength,structure of osteopenic femurs and BMD.Our findings may provide a new strategy for the prevention and treatment of osteoporosis,especially for osteoporotic fractures.

Objective To analyze the clinicopathologic characteristics,treatment and prognostic factors in malignant transformation of mature cystic teratoma(MCT)of ovary.Methods The clinical data of 44 patients with MCT from January 1961 to June 2009 were reviewed.Results The median age of the 44 patients was 48 years(range,16-84 years).Mean tumor size was(16 ±6)cm.Thirty-two cases were diagnosed squamous cell carcinoma(73％,32/44),and 5 of them with the elevated level of serumal squamous cell antigen(SCC-Ag).Three of 37 cases(8％,3/37)were identified with malignant transformation in image examinations.Rapid frozen section examination and multiple-location biopsy were performed in 8 cases,and 5 of them were detected with malignant diseases.Twenty-two patients with disease confined within the unilateral ovary(10 with intact capsule,and 12 with ruptured capsule).Diseases extended extra ovaries in the others 22 patients.The median cumulative overall survivals were 126 and 10 months,respectively.The difference between the two groups was significant(P ＜ 0.01).Twenty-seven patients had no residual tumor after primary surgery.The median cumulative overall survivals between the patients with and without residual tumor were 10 and 84 months respectively,and there were significant difference between two groups(P ＜ 0.01).Seven selected patients with malignant disease confined within unilateral ovary underwent fertility-sparing surgery,and 2 cases of them had successful pregnancies and delivery,while other 4 cases with ruptured capsule recurred.Conclusions The most common pathology type of malignant transformation in mature cystic teratoma of the ovary is squamous cell carcinoma.Comprehensive pre-operation image examination and tumor marker level detection might be of great help in diagnosis.Tumor extension extraovary and residual tumor after surgery are the most significant poor prognostic factors.Early stage patient with ruptured capsule should be very discreet to choose fertility-sparing surgery.

Objective:This study analyzes the clinicopathological characteristics, pathological diagnosis, treatment, and prognosis of ovarian small cell carcinoma (SCCO). Methods:The medical records of SCCO patients in the Cancer Hospital of Peking Union Medical College between 2005 and 2012 were reviewed. Results:The mean age of patients was 43. 75 years old (ranging from 17 to 57), two cases were postmenopausal, and one case was less than 30 years old. Twenty-one patients had FIGO stageⅠ, whereas three cases had stageⅢto Ⅳ. Three cases were classified as pulmonary type. Forty-three patients received cytoreductive surgery, and one underwent fertili ty-conserving surgery. All patients were postoperatively treated with platinum-based chemotherapy, whereas three cases received Paclitaxel plus Carboplatin. No patient received adjuvant radiotherapy. One patient died nine months after the initial diagnosis, and one died 12 months after the initial diagnosis. The other two cases remain alive with no evidence of recurrence after follow up at 7 and 30 months after diagnosis. Conclusion:SCCO is a rare ovarian tumor with high malignancy potential and thus has poor prognosis. The clinical manifestations of SCCO resemble those of epithelial ovarian cancer. Immunohistochemistry can be used for differential diagnosis. The standard SCCO treatments are cytoreductive surgery and adjuvant platinum-based chemotherapy. The optimal chemotherapy regimen requires further research.