Preliminary investigation of intramural lateral spread distance in pull-through conformal resection of low rectal cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the intramural lateral spread distance in low rectal cancer in order to provide basis for safety lateral resection margin of pull-through conformal resection (PTCR).</p><p><b>METHODS</b>The patients with low rectal cancer who received low anterior resection or abdominal-perineal resection in Changhai Hospital from December 2015 to March 2016 were enrolled and Surgical specimens were collected. After the specimens were fixed in 10% formaldehyde for 24 hours, a piece of tissue that was 1.5 cm in length and 0.5 cm in width from the edge of tumor was cut. The tissue was obtained in the direction of 3, 5, 7 and 9 o'clock clockwise. The distance of intramural lateral spread was measured in the specimens and the risk factors were analyzed.</p><p><b>RESULTS</b>A total of 83 specimens were collected and the overall proportion of intramural lateral spread was 71.1%(59/83). The rate of lateral spread from 3 to 9 o'clock was 34.9%(29/83), 26.5%(22/83), 32.5%(27/83) and 37.3%(31/83) respectively, and the difference was not statistically significant(χ=2.444 9, P=0.485 3). The median distance of lateral spread in each direction was all 0 mm and the quartile range was 1 mm, 0.5 mm, 0.55 mm and 1 mm respectively. The 5th percentile (P5) of each direction was all 0 mm and the 95th percentile(P95) of each direction was 2.5 mm, 1.6 mm, 2.6 mm, 2.5 mm, respectively and the difference was not statistically significant either(χ=5.331 0, P=0.148 9). The rate of lateral spread of T1, T2, T3 and T4 was 0/4, 58.3%(14/24), 83.0%(44/53) and 1/2 respectively, and there was significant difference(P=0.005 0). The multivariate analysis indicated that T stage (P=0.002 2, OR=3.741, 95% CI: 1.606-8.716) was the risk factor of intramural lateral spread.</p><p><b>CONCLUSIONS</b>The intramural lateral spread does exist in low rectal cancer and T stage is the risk factor of lateral spread. The lateral resection margin should be 5 mm from the tumor edge at least when PTCR is performed.</p>

Consistency analysis of gastric cancer HER-2 expression among endoscopic biopsy and radical operation specimens / 中国肿瘤临床

Objective:To analyze the consistency of HER-2 expression among endoscopic biopsy and radical operation specimens of gastric adenocarcinoma and to investigate the clinical application value of HER-2 detection in trastuzumab treated patients. Methods:From March 2013 to February 2014, 167 patients from Shanghai Changhai Hospital were diagnosed with gastric adenocarcinoma using endoscopic biopsy specimens. The corresponding surgical specimens were collected for pathological analysis. The relevant clinical and pathological data were collected. HER-2 protein expression of endoscopic biopsy specimens was detected by immunohistochemistry (IHC). HER-2 protein expression and gene amplification status of the corresponding tumor resection specimens were detected by IHC and fluorescence in situ hybridization (FISH). Results were analyzed for clinicopathological characteristics. Results:Among the 167 cas-es, 18 cases (10.8%) were HER-2 positive, including 10 cases showing IHC3+and 8 cases showing IHC2+with FISH positive. The consis-tency rate result among endoscopic biopsy and surgical operation specimens was 82%. Excluding the cases showing IHC2+, the true positive rate and the true negative rate were 73.3%and 97.0%, respectively. Conclusion:HER-2 detection of endoscopic biopsy speci-men by IHC shows great predictive value. The main reason for the difference of surgery and biopsy specimens is the heterogeneity of tumor expression. Increasing the number of specimens and combined testing with FISH are important methods to reduce misjudge-ment.

Construction of tissue engineering adult cardiac myocytes / 中国组织工程研究

BACKGROUND:Construction of tissue engineering adult cardiac myocytes has been a new research hotspot in cardiovascular fields. OBJECTIVE:To explore a simple, fast method for the separation of adult rat cardiomyocytes, and preliminarily explore the construction methods of tissue engineering adult cardiac myocytes. METHODS:Segmented enzymatic digestion method was used to isolate cardiac myocytes from adult rats. Subsequently, cardiac myocytes were transfected with adenovirus and liposome-mediated red fluorescent protein gene. Construction efficiency of tissue engineering cells was qualified using inverted fluorescence microscopy and flow cytometry. Final y, cardiac myocytes were transfected with adenovirus-mediated hypoxia-inducible factor-1a, and the expression of hypoxia-inducible factor-1a was examined by western blot analysis. RESULTS AND CONCLUSION:A lot of cardiac myocytes were col ected using the segmented enzyme digestion method. Flow cytometric analysis showed that the survival of adult cardiomyocytes was (87.03±0.70)%. Compared with liposomal transfection (transfection efficiency was 0), adenovirus infection efficiency was (70.31± 1.39)%, and the cells expressed red fluorescence under fluorescence microscope. After 4 days of adenovirus infection, transfected cells expressed hypoxia-inducible factor-1a protein. These results showed that segmented enzyme digestion is a fast way to isolate adult cardiac myocytes, and recombinant adenovirus vector is a good vector to transfect cardiac myocytes.

The Sensitivity of Gene Mutant Related to Gastrointestinal Stromal Tumor to Gleevec / 中国肿瘤临床

Objective: To explore the sensitivity of Kit or PDGFRA mutants related to gastrointestinal stromal tumor (GIST) to Gleevec.Methods: The recombinant plasmids of KIT Del559-560, KIT Ins IPYD579, PDGFRA D842V and PDG-FRA L839P gene mutants were transiently transformed into the CHO cells by liposome methods.Western blot was used to detect the expression of the related protein and their phosphorylated forms after the cells were incubated with Gleevec for 90 min.At 72 hours after incubation with Gleevec, MTT was used to detect cell proliferation.Results: Western blot results showed that Gleevec at 0.1 μM can notably reduce phosphorylation of KIT Del559-560.Gleevec at 1μM completely blocked phosphorylation of KIT Ins IPYD579 and PDGFRA L839P, but did not affect PDGFRA D842V phosphorylation.MTT analy-sis indicated that growth of CHOPDGFRA L839P was inhibited by Gleevec at 1μM, however, CHOPDGFRA D842V was re-sistant to Gleevec at 5 μM.Conclusion: Gleevec can decrease the expression of phosphorylated protein CHOPDGFRA L839P and CHOKIT Ins IPYD579, and can remarkably inhibit the proliferation of cells containing PDGFRA L839P mutant.

Expression and clinical significance of c-kit oncogene in gastrointestinal stromal tumors / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathological and prognostic significance of the c-kit protein in gastrointestinal stromal tumor (GIST).</p><p><b>METHODS</b>Paraffin embedded materials from 53 benign GISTs, 13 potentially malignant and 55 malignant cases were analysed for c-kit expression by immunohistochemical method, while using leiomyomas and schwannomas as controls. Positive signals were shown in cytoplasma and cell membrane.</p><p><b>RESULTS</b>Of 122 GISTs, 118 (97%) were positive for c-kit. Localization of positive signals was accurate. The rate of c-kit protein in benign, potentially malignant and malignant cases was 98% (53/54), 93% (12/13), 96% (53/55) respectively. Compared with benign GIST, the positivity of c-kit in metastasis or recurrent cases decreased, but c-kit protein expression rate was not significantly different between the three patterns of GIST (chi(2) = 1.167, P > 0.05). Leiomyomas and schwannomas were typically c-kit negative.</p><p><b>CONCLUSION</b>As a sensitive and specific marker of GIST, c-kit seems to be a useful antibody in the diagnosis and differential diagnosis of GIST, but it may not be used as a prognostic index.</p>

Expression of human giant larvae-1 in ovarian cancer and its clinical significance / 第二军医大学学报

Objective To study the expression of human giant larvae-1 (Hugl-1) in ovarian carcinoma and its clinical significance.Methods Hugl-1 mRNA expression in 31 ovarian cancer and the corresponding adjacent tissues was examined by real time fluorescence quantitative RT-PCR and in situ hybridization.Moreover,analysis was done while taking into consideration of the clinicopathologic parameters of ovarian cancer.Results The expression of Hugl-1 mRNA in ovarian cancer tissue was significantly higher than that in the corresponding adjacent tissues ([0.051 ? 0.029]vs [0.026 ? 0.043],P

Construction of recombinant plasmids with mutant C-kit cDNA in gastrointestinal stromal tumor / 第二军医大学学报

Objective:To construct the recombinant eukaryotic expression vector plasmids with mutant C-kit cDNA and to study the effect of the mutant C-kit gene on cell proliferation and cell cycle in gastrointestinal stromal tumor (GIST). Methods: Wild-type C-kit cDNA was cloned from human embryonic brain tissue by RT-PCR technique.Site-directed mutagenesis of the wild type C-kit cDNA was performed according to the C-kit mutations we cloned. Recombinant plasmids were stably transfected into human embryonic kidney cell line and the cells expressing mutant C-kit were selected by special cell culture medium containing G418.Expressions of C-kit protein of the transfectants were detected by Western blot.Cell proliferation and cell cycle of the transfectants were detected by MTT clolorimetic assay and flow cytometry,respectively.Whether HEK cell with mutated C-kit cDNA could grow autonomously in nude mice or not was also detected. pcDNA3 vector transfected and recombinant plasmids with wild-type C-kit cDNA transfected HEK cell were used as the control groups. Results: The mutant C-kit cDNA was obtained by site-directed mutagenesis of the wild type C-kit cDNA. Compared with the 2 control groups ,the growth rate and proliferative activity of the HEK cells with mutant C-kit cDNA were increased significantly.The analysis of cell cycle showed that more HEK cells with mutanted C-kit cDNA remained in proliferation phase (S+G 2-M)than the groups without mutated C-kit cDNA.HEK cells with the mutated C-kit also grew autonomously in nude mice.Conclusion: Mutation of C-kit gene can increase proliferation of human cells,causing malignant transformation of human normal cells,which may play an important role in the malignant transformation of GIST.

A method for detection of HER2 gene amplification in breast cancer tissues:chromogenic in situ hybridization method and its modification / 第二军医大学学报

Objective:To apply chromogenic in situ hybridization(CISH)for detection of HER2 gene amplification in breast cancer tissues and to discuss some modifications of the CISH method.Methods:HER2 gene amplification was detected by CISH in 60 breast cancer specimens with an immunohistochemical score over 2+.The correlation between the results of IHC and CISH was analyzed.Our experience in CISH manipulation was summarized and optimization to CISH was discussed.Results:CISH identified gene amplification in 91%(40/44)specimens with an IHC score of 3+ and in 50%(8/16)specimens with an IHC score of 2+.The total concordance rate between IHC and CISH was 80%(48/60,P