Expressions of large tumor suppressor kinase 1 and large tumor suppressor kinase 2 in gastric cancer and their significances / 肿瘤研究与临床

Objective To investigate the expressions of large tumor suppressor kinase 1 (LATS1) and large tumor suppressor kinase 2 (LATS2) proteins in gastric cancer tissues, and to explore the correlation between expressions of LATS1 and LATS2 proteins and the occurrence and development of gastric cancer. Methods A total of 93 gastric cancer paraffin tissues and the corresponding adjacent gastric normal mucosa in the Department of Pathology in Baotou Cancer Hospital from September 2008 to June 2010 were collected. The immunohistochemistry was used to detect the expressions of LATS1 and LATS2 proteins in gastric cancer and adjacent normal tissues. The differences of the expressions of LATS1 and LATS2 proteins in gastric cancer and adjacent normal tissues were compared by usingχ2 test. The relationship between the expressions of LATS1 and LATS2 proteins and the clinicopathological features was also analyzed. Results In gastric cancer tissues, LATS1 was negatively expressed in 54 cases (58.1％), weakly positive expressed in 15 cases (16.1％), moderately positive expressed in 16 cases (17.2％), and strongly positive expressed in 8 cases (8.6％);in adjacent normal tissues, LATS1 was negatively expressed in 17 cases (18.3％), weakly positive expressed in 16 cases (17.2％), moderately positive expressed in 31 cases (33.3％), and strongly positive expressed in 29 cases (31.2％). The positive expression rate of LATS1 in gastric cancer tissues was lower than that in adjacent normal tissues, and the difference was statistically significant (χ2=37.460, P<0.01). In gastric cancer tissues, LATS2 was negatively expressed in 28 cases (30.1％), weakly positive expressed in 17 cases (18.3％), moderately positive expressed in 33 cases (35.5％), strongly positive expressed in 15 cases (16.1％);in adjacent normal tissues, LATS2 was negatively expressed in 5 cases (5.4％), weakly positive expressed in 7 cases (7.5％), moderately positive expressed in 32 cases (34.4％), strongly positive expressed in 49 cases (52.7％). The positive expression rate of LATS2 in gastric cancer tissues was lower than that in adjacent normal tissues, and the difference was statistically significant (χ2=38.275, P<0.01). The expressions of LATS1 and LATS2 were not related to patients'age, gender, lymph node metastasis, degree of differentiation and tumor diameter (all P>0.05). Conclusion LATS1 and LATS2 proteins may be involved in the occurrence of gastric cancer and have the inhibiting effect on the occurrence and development of gastric cancer.

Identification of epitope recognized by mAb 15 A11 sepecific against cartilage oli-gomeric matrix protein / 中国免疫学杂志

Objective:To identify the epitope of mAb15A11 which is specific against RA associated autoantigen cartilage oligomeric matrix protein ( COMP ).Methods: A filamentous phage library displaying random linear dodecapeptides was used to mapping the epitope of mAb15A11.After three rounds of screenings,40 phage clones were selected at random and sequenced.The specificity of phages was confirmed by enzyme immunoassays.Homology search by ClustalW2 and structure analysis by PyMol to identified the epitope amino acid sequence.Western blot analysis of COMP and ELISA analysis of COMP-derived peptides were used to confirm epitope′s characterization.Results: After repeated screenings using bio-panning method, 2 clones were identified, which interacted specifically with mAb 15A11.Homology search did not find succession consensus sequence within COMP molecular,which indicated that the epitope was not linear.PyMol Structure analysis identified the rationality of conformational epitope.Western blot analysis and ELISA of EDTA-treated COMP further prove an conformational structure of the epitope recognized by mAb 15A11.ELISA analysis of COMP-derived peptides demonstrated both disulfide bonds between 229 C-243 C and 237 C-253 C and every epitope amino acid of 232 G,238 H,240 H,241 A,244 V,247 R and 251 R were essential to the binding of mAb 15A11 with COMP.Conclusion: In this study, the potential B cell antigentic epitopes of mAb 15A11 was identified by phage display library.The epitope amino acids sequence and char-acterization were also recognized.It may have important theoretical value for the study of reaction mechanism of COMP antibody and antigen and may also show application significance in the detection of rheumatoid arthritis.

Analysis of gene structure, cloning and expression of cyp51 from Ustilago maydis / 生物工程学报

The cyp51 primers and two pairs of mutant primers which removed different transmembrane region were designed based on Ustilago maydis cyp51 gene structure analysis. The full cyp51 DNA fragment as well as mutant cyp51 genes were amplified and cloned by using the total DNA from Ustilago maydis as template, then subcloned into different expression vectors. The recombinant expression plasmids were transformed into Escherichia coli BL21 (DE3), BL21 (DE3) pLysS and Rosetta (DE3) respectively. A series of experiments leads to the finding that only pET32-YH-35 could be highly expressed at the optimal condition of 30 degrees C induced with 0.5 mmol/L IPTG The expressed protein (CYP51) showed biological activity by spectra analysis of the protein binding to 4 standard fungicides and to 14 XF-synthetic fungicide compounds, and only one XF-synthetic fungicide compound (XF-113) was similar to standard fungicides in binding constant. This compound is promising to be a new effective antifungal drug. These results will facilitate the further study on the mechanism of pathogenic fungi CYP51 and pesticide molecules, and will provide a new idea for efficient design and development of new anti-fungal drugs.

Construction of recombinant human smad 7 adenoviral vector and its expression in vitro / 生物医学工程学杂志

The recombinant human Smad7 adenoviral vector was constructed by direct DNA cloning protocol and then transfected into 293 cells for virus packaging. After amplification and purification, the recombinant adenovirus was used to infect the keloid fibroblasts. The Smad7 mRNA transcription of the infected cells was detected by RT-PCR. The recombinant Adeno-Smad7 was correctly constructed and confirmed by both restriction analysis and PCR analysis. RT-PCR showed the over expression of adenovirus mediated Smad7 mRNA in keloid cells. These results demonstrated that the recombinant Smad7 adenoviral vector can be expressed in cultured cells in vitro, and it may provide a new therapeutic strategy for keloid gene therapy.

Perinatal Outcomes in Pregnancies with Toxoplasma, Cytomegalovirus and Herpes Simplex Type Ⅱ Detected in Maternal Blood / 中华围产医学杂志

Objective To study the perinatal outcomes in pregnant women with pathogens of toxoplasma,cytomegalovirus and herpes simplex type Ⅱ in maternal blood.Methods 1 141 preg- nant women were screened for toxoplasma,eytomegalovirus and herpes simplex type Ⅱ by PCR in maternal blood during 1994.2～1997.3.Some placental tissues and/or umbilical vein blood were also detected.Results Toxoplasma,cytomegalovirus,or herpes simplex type Ⅱ were positive in 97 cases(8.50%).All cases,except that 25 cases did not deliver in our hospital,were followed up to 42 days postpartum.Fifty-two cases got good perinatal outcomes,while 20 cases had poor perinatal outcomes.17 placentas were detected in patients with good perinatal outcomes,while 15 placentas were detected in patients with poor perinatal outcomes.The positive rates of toxoplasma,cytomega- lorirus and herpes simpley type Ⅱ in the two groups were 17.65%(3/17)and 60.00%(9/15)re- spectively(P

Stability of ferulic acid and its existing form in Ligusticum chuanxiong and Angelica sinensis / 中草药

Object To investigate the isomer transition reaction of ferulic acid and the existing form of ferulic acid in Ligusticum chuanxiong Hort. and Angelica sinensis (Oliv.) Diels. Methods HPLC and HPLC-MS were used to confirm the isomer transition of ferulic acid. Results The trans-ferulic acid was translated into cis-ferulic acid partly when its solution was deposited under some routine conditions,and the trans- and cis-ferulic acids could coexist in the Chinese medicinal materials,L. chuanxiong and A. sinensis. Conclusion The trans-ferulic acid is stable thermodynamicly,but could be translated into cis-ferulic acid partly and slowly.