Objective To investigate the expressions of cycloxygenase-2 (COX-2 ),vascular endothelial growth factor (VEGF),matrix metalloproteinase (MMP2 )and micro-vessel density (MVD)in papillary thyroid carcinoma (PTC)and the relationship between their expressions and clinicopathological features,so as to evaluate the malignancy and prognosis of PTC.Methods The expressions of COX-2,MMP2 and VEGF and MVD count in 32 cases of PTC and 18 cases of normal thyroid tissues were detected by immunohistochemical staining.Their relationship with clinicopathological characteristics was analyzed.Results ① The expression of COX-2,MMP2, VEGF and MVD in PTC was as follows:M(Q 3 -Q 1 )=5(1),M(Q 3 -Q 1 )=5.5(2),M(Q 3 -Q 1 )=5.5(1)and M (Q 3 -Q 1 )= 28 (5.75 ),respectively.It differed significantly from that in control group (P < 0.05 ).② The expression of COX-2 in PTC was related to the sex and age of patients,clinical stage and lymph node metastasis (P <0.05).The expression of VEGF in PTC was related to the sex and age of patients,tumor size,clinical stage and lymph node metastasis (P <0.05).The expression of MMP2 and MVD count in PTC were related to the age of patients,tumor size,clinical stage and lymph node metastasis (P <0.05 ).③ The expression of COX-2,MMP2, VEGF and MVD in PTC with invasion of thyroid capsules was as follows:M(Q 3 -Q 1 =6(2),M(Q 3 -Q 1 )=6.5 (2),M(Q 3 -Q 1 )=6(1.75)and M(Q 3 -Q 1 =30(7.75).The expression of these indexes in negative group was:M (Q 3 -Q 1 )=5(1.75),M(Q 3 -Q 1 )=5(1.75),M(Q 3 -Q 1 )=5 (1.75 )and M (Q 3 -Q 1 )=26.5 (4).There was a significant difference between the two groups (P < 0.05 ).④ There was a close positive correlation between the expressions of VEGF,COX-2,MMP2 and MVD in PTC (r >0.5).Conclusion The high expressions of COX-2, VEGF and MMP2 in thyroid tissues may result in the occurrence of PTC and lymph node metastasis,which is related to the regulation of tumor new vessels.Detecting these expressions are of value in evaluating the malignancy and prognosis of PTC.

Objective To detect the expressions of proliferating cell nuclear antigen (PCNA), C-erbB-2, p53, estrogen receptor (ER) and progesterone receptor (PR) in order to explore the proper margin for breast conservative surgery on Chinese women. Methods We collected 40 resection specimens from breast cancer patients who had received radical surgery. Then we divided each specimen into primary tumor group and hyperplasia and gene expression characteristics of PCNA, c-erbB-2, p53, ER and PR were measured by pathological and immunohistochemical assay in the five groups. Results With the further distance from the primary tumor, the proportions of high-risk disease and positive gene expressions of PCNA, c-erbB-2 and p53 in paracarcinoma tissues gradually decreased (P<0.05). Higher risk and more positive expressions were related to paracarcinoma was no correlation of PCNA, ER and PR expressions adjacent to breast cancer with tissue differentiation and lymph node metastasis (P>0.05). a safe and appropriater region for breast conservative surgery.

Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropylβ-D-thiogalactopyranoside (IPTG) at 37 ℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation. The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results:The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21,and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14 000.Two bands (14 000 and 28 000)were seen in the SDS-PAGE picture,which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg · L-1 , respectively, and the recovery rate was about 73%. Conclusion:The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully,and the recombinant CBD-BMP2 is expressed and refolded efficiently. The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.