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 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 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 investigate the prevention of stroke-associated pneumonia (SAP) with Ramipril.Methods 404 acute stroke patients with primary hypertension were randomized into two groups.Group A received ramipril 2.5 mg once daily orally,and then were titrated up to 5 mg/d after a week.Group B received Candesartan 4 mg once daily orally,and then were titrated up to 8 mg/d after a week.The treatment course was 28 days.All patients received examination of Standardized Swallowing Assessment (SSA).The amelioration of swallow function and incidence of SAP after treatment were compared between the two groups.Results There were 10 patients suffering from SAP in group A and 21 patients in group B.The incidence of SAP was lower in group A(5.0％)than in group B(10.4％)(P＜0.05).The positive rate of SSA was 87.1 ％ in patients with SAP,higher than the total rate (47.5％).Dysphagia was improved in both groups after treatment.The rate of dysphagia improvement in group A and group B was 81.4％ and 66.3％ respectively.There was significant statistics difference in the positive case of SSA between two group (P＜ 0.05).Conclusions Orally treatment with ramipril is effective to prevent stroke-associated pneumonia and toimprove swallow function.

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.

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.

Objective:To analyze the learning effect of laparoscopic radical cystectomy(LRC)+ modified ileal conduit(MIC).Methods:From 2014 to 2019, 42 patients underwent MIC and their clinical data was retrospectively analyzed. 34 operations were performed by surgeon 1 and 8 operations by surgeon 2. We divided the 34 patients of surgeon 1 into three groups according to their surgical sequence (group A, 1st to 12th; group B, 13th to 23th; group C, 24 th to 34 th), the 8 cases of surgeon 2 was regarded as group D. The history of abdomen surgery in the 4 groups were 0, 1, 4, 3 cases, respectively ( P<0.05). There was no significant difference of the other baseline characteristics, such as age, BMI, American Society of Anesthesiologists. Then we compared several variables between the 4 groups like operation time, time of ileal conduit construction, blood loss, complication rate, lymph node yield, surgical margin, etc. The key steps of the MIC included isolating terminal ileum when the mesentery was transilluminated, performing end-to-end reflux ureterointestinal anastomosis after the efferent loop was fixed, closing the rent of the retroperitoneum. Results:All operations were performed intracorporeally with no transition to open surgery. The operative time for group A, B, C were 330.0(320.0, 360.0)min, 300.0(250.0, 308.0)min, 270.0(216.0, 324.0)min, respectively ( P =0.010). The time of ileal conduit construction of the 3 groups were 136.5(131.3, 147.5)min, 92.0(79.0, 119.0)min, 79.0(72.0, 115.0)min, respectively ( P <0.001). In addition, the difference of the two variables above between A and B, A and C groups separately reached statistical significance ( P<0.05), while the difference between B and C groups did not ( P>0.05). Other variables, such as blood loss, complication rate, lymph node yield, surgical margin, between the 3 groups reached no statistical significance ( P>0.05). The operative time of group D was 420.0(350.0, 450.0)min, and it reached statistical significance ( P<0.05) when compared with group A. There were no significant differences in other variables, such as blood loss, complication rate, lymph node yield, surgical margin, among the 2 groups ( P>0.05). Conclusions:The learning effect of LRC+ MIC was obvious. When surgeon volume increased, the operative time decreased significantly. Variables like estimated blood loss and complication rate of the 2 surgeons did not reached significant difference, which indicated reproductivity and safety of this procedure.

Objective To investigate the feasibility, efficacy, safety and economy of secondary splenic pedicle trisection method in removing schistosoma cirrhosis caused the splenic function. Methods Thirty patients receiving spleen secondary structure amputation between July 2014 and September 2016 were analyzed. Results Laparoscopic splenectomy with secondary splenic pedicle transaction was successfully performed in 28 patients, whereas two Endo-GIAs were used in 2 patients. The average of operation time was (80 ± 20) min, and operative blood loss was (320 ± 10) ml. The drainage of the splenic fossa was removed (3- 4) days after operation.Postoperative hospital stay was (10.8 ± 1.2) days after operaions. No massive hemorrhage, pancreatic leakage, secondary infection, serious complications such as abscess under diaphragm and recent complication such as infection of incision occurred postoperatively. Platelet of all patients recovered in 4 days postoperatively, and patients with platelet>400 × 109/L was given oral aspirin enteric-coated metformin hydrochloride. All patients were followed up for 6 months postoperatively, and no intestinal obstruction, portal vein thrombosis and other long-term complications occurred in all patients. Conclusions The amputation of secondary structures of the spleen in laparoscopic splenectomy to remove schistosoma cirrhosis caused the splenic function is safe. It could shorten the length of hospital stay and reduce the medical cost. It is a valuable method for clinical promotion.