Correlation of clinicopathologic features and postoperative recurrence or metastasis in solid-pseudopapillary neoplasms of the pancreas / 临床与实验病理学杂志

Purpose To explore the correlation between the clinicopathological features and postoperative recurrence or metastasis of solid-pseudopapillary neoplasms of the pancreas (SPN).Method The clinicopathological characteristics of 73 SPNs were summarized,the patients' prognosis were followed up by telephone and then the correlation of clinicopathological characteristics and recurrence or metastasis was analyzed.Results 57 female patients and 16 male patients were included in this study.The age range was 7 to 68 years old with the average of 28 and median age of 27.The mean diameter of the tumors was 6.47 cm (range 0.31～ 14 cm).30 cases of tumors were located in the head of pancreas,9 in the body of pancreas,and 33 in the tail of pancreas.One case was a multiple lesion simultaneously located in the body and the tail.All patients were followed up by telephone for mean time 34.8 months with the range of 12 ～ 99 months.Necrosis,calcification,cholesterol crystal,foamy histiocytes,nuclear atypia,pancreatic parenchymal invasion,and perineural invasion had no statistical significance between non-recurrent/metastatic group and recurrent/metastatic group.However,there was significant difference for extra-pancreatic invasion,angiovascular invasion and Ki-67 proliferation index between non-recurrent/metastatic group and recurrent /metastatic group.Conclusion Extra-pancreatic invasion,angiovascular invasion and Ki-67 proliferation index ≥ 4％ have reference significance for predicting recurrence or metastasis of SPN.

Construction of chimeric E3s expression plasmids targeting oncoprotein ras / 中国医学科学院学报

<p><b>OBJECTIVE</b>To construct certain chimeric E3s expression plasmids targetting oncoprotein Ras by harnessing the theory of protein knockdown.</p><p><b>METHODS</b>We chose the binding domain of Raf-1, PI3K, RalGDS, and the function domain of F-Box as well as the U-Box to construct the plasmids. Then used the double enzyme, PCR, and sequence to test the validity and integrity of the cloned nucleotide fragments. The expression efficiency of the plasmids in eukaryotic cells was detected by Western blot analysis.</p><p><b>RESULTS</b>Five of 6 plasmids in this study expressed the corresponding fusion proteins in HEK293T cells, and (RBD+CRD)(Raf-1)- U-Box-pcDNA3.1 can knocked down the protein level of Ras in PANC-1 cells.</p><p><b>CONCLUSIONS</b>We successfully constructed the chimeric E3 expression plasmids, which provides a solid basis for further research on protein knockdown.</p>

Construction of a miR-23a-27a cluster expression plasmid: a preliminary study of its function / 中华病理学杂志

<p><b>OBJECTIVE</b>To construct a miR-23a-27a cluster expression plasmid and to explore the target genes and function of the cluster.</p><p><b>METHODS</b>The pre-miR-23a-27a-pcDNA3.1, pre-miR-23a and pre-miR-27a plasmids were cloned by molecular biology method, and their expression efficiency was tested by dual luciferase reporter gene assay and real-time PCR. Several possible target genes of miR-23a and miR-27a were chosen using softwares and further tested by dual luciferase reporter gene assay. Finally, the function of miR-27a was analyzed in MCF-7 cell by Western blot and real-time PCR.</p><p><b>RESULTS</b>miR-23a and miR-27a were transcribed from pre-miR-23a-27a-pcDNA3.1, pre-miR-23a and pre-miR-27a plasmids in HEK293T cells, and both influenced the MRE of Sprouty2 gene in pRL-TK vector, and only miR-27a influenced the 3'-untranslated regions (UTR) full length of Sprouty2 gene while miR-27a did not influence the 3'-UTR of Sprouty2 gene with the sited-mutation in the MRE. The protein expression level of Sprouty2 gene was altered after transfection of pre-miR-27a-pcDNA3.1 plasmid while the RNA level remained unchanged.</p><p><b>CONCLUSION</b>Sprouty2 may be the functional target gene of miR-27a, and the construction of plasmids in the study may provide a fundamental basis for the further functional investigation of miR-23a and miR-27a.</p>

Significance of chemokine receptor CXCR3 expression in breast cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of CXCR3 and its association with clinicopathologic features in breast carcinoma.</p><p><b>METHODS</b>The expression level of CXCR3 in 18 samples of breast cancer and corresponding normal tissues was investigated using reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time RT-PCR analysis. Immunohistochemistry was carried out to examine the expression of CXCR3 in 80 breast cancers, 20 fibroadenomas and 15 normal breast tissues.</p><p><b>RESULTS</b>(1) RT-PCR and real-time RT-PCR analysis showed a higher level of CXCR3 in breast cancer tissues than that in the corresponding normal breast tissues (P < 0.05). (2) Immunohistochemistry analysis showed that the positive rate of CXCR3 in breast cancer tissues was significantly higher than that in fibroadenomas and the normal breast tissues (P < 0.05). The expression level of CXCR3 in the lymph node-positive group was higher than that in the lymph node-negative group (P < 0.05). The expression of CXCR3 was positively correlated with the number of lymph nodes involved by metastasis, tumor size and pTNM tumor stage (P < 0.05).</p><p><b>CONCLUSIONS</b>Chemokine receptor CXCR3 was up-regulated in breast cancer, and was associated with the progression of breast cancer. CXCR3 might be a novel molecular marker to predict lymph node metastasis and prognosis of breast cancer.</p>

Expression of Twist, E-cadherin and N-cadherin in breast carcinoma and their clinical significance / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of Twist, E-cadherin and N-cadherin in breast carcinoma tissue and to analyse their effects on the breast carcinoma differentiation, size, infiltration and metastasis of the breast carcinoma.</p><p><b>METHODS</b>The expression of Twist, E-cadherin and N-cadherin in 56 cases of breast invasive ductal carcinoma, 38 cases of invasive lobular carcinoma, 41 cases of carcinoma in situ and 10 cases of normal breast tissue was detected using immunohistochemistry.</p><p><b>RESULTS</b>(1) The expression rate of Twist in three types of breast carcinoma was 46.4% (26/56), 79.0% (30/38) and 26.8% (11/41) respectively, and the expression of Twist in invasive lobular carcinoma was significantly higher than that in invasive ductal carcinoma and carcinoma in situ (P = 0.002, P = 0.000). The expression rate of E-cadherin in three types of breast carcinoma was 78.6% (44/56), 29.0% (11/38) and 80.5% (33/41) respectively, and the expression of E-cadherin in invasive ductal carcinoma and carcinoma in situ was significantly higher than that in invasive lobular carcinoma (P = 0.000, P = 0.000). The expression rate of N-cadherin in three types of breast carcinoma was 53.6% (30/56), 68.4% (26/38) and 31.7% (13/41) respectively, and the expression of N-cadherin in invasive ductal carcinoma and invasive lobular carcinoma was significantly higher than that in carcinoma in situ (P = 0.033, P = 0.001). (2) In all the 135 cases, the expression of Twist was not correlated with that of E-cadherin (P = 0.005, Spearman correlation coefficient = -0.239), however, there was a positive correlation between the expression of Twist and N-cadherin and statistically significant(P = 0.000, Spearman correlation coefficient = 0.319). (3) In the invasive ductal carcinoma, the expression of N-cadherin in poorly-differentiated carcinoma was significantly higher than that of the moderately-or well-differentiated ones (P = 0.004). (4) In the invasive lobular carcinoma, the expression of Twist in cases with lymph node metastasis was significantly higher than that of cases without metastasis (P = 0.037).</p><p><b>CONCLUSIONS</b>Twist, E-cadherin and N-cadherin have different expression patterns in the three kinds of breast carcinoma. The positive expression of Twist was correlated to lymph node metastasis in invasive lobular carcinoma and the positive expression of N-cadherin was correlated to cell the tissue differentiation in invasive ductal carcinoma. Detection of the expression of these biomarkers may provide a valuable reference for the study of breast carcinoma progression, metastasis and for the judgment of the biological behavior of the carcinoma.</p>

Utilization of targeting specific proteins for ubiquitination and degradation / 中国医学科学院学报

Ubiquitin-proteasome pathway (UPP) can degrade specific proteins in eukaryotic cells. Targeting specific proteins for ubiquitination and degradation technique, based the theory of UPP and using certain chimeric E3s or protein-targeting chimeric molecules, can specifically knock down the targeted proteins. It is a promising new method for exploring the functions of genes or proteins and, together with gene-silencing technique, may play an important role in basic and clinical research.

Effect of survivin antisense mRNA transfection on the growth and chemotherapy sensitivity of lymphoma cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the effect of transfecting survivin antisense mRNA on growth and chemotherapy sensitivity of lymphoma cells.</p><p><b>METHODS</b>Eukaryotic expression plasmid pcDNA3. 1-antisense (As) survivin was constructed and transfected into Jurkat T lymphoblastic lymphoma cell lines with high expression survivin mRNA by use of lipofectmine gene transfer technique. Expression of survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical and Western blot. The effect of transfecting survivin antisense mRNA on the growth of Jurkat cell lines was monitored by population doubling time (PDT) and Apoptotic indexes (AI). The morphologic features were observed in transfected cells by light and electric microscopes. MTT assay was used to analyze the response of transfected cells to CTX and MTX.</p><p><b>RESULTS</b>Compared with the control cells, the expression of survivin mRNA and protein were reduced after transfected pcDNA3. 1-Assurvivin 48 h, 5 w and 6 w, PDT (52 h) was prolonged. Apoptotic indexes were higher in transfected antisense survivin mRNA cells [20.2% (48 h)], 6.2% (5 w) and 6.8% (6 w) than control ones [2.1%, 1.3% (48 h)] and [1.3% (5 w) and 1.0% (6 w)]. The cells grow slowly and the dead cells increase and some swelling and apoptotic cells were observed in transfected pcDNA3. 1-Assurvivin groups by invert, light and electric microscopes. The Jurkat cell line of transfected pcDNA3. 1-Assurvivin had higher sensitivity to CTX and MTX. The rate of inhibition was higher in transfected group. There is a significant difference between the transfected group and untransfected one, P < 0.05.</p><p><b>CONCLUSIONS</b>The result indicated that survivin gene was very important for growth of Jurkat cells. To inhibit the expression of survivin will be significant in therapy of T lymphoblastic lymphoma. Survivin gene might be a target of therapy.</p>