Therapeutic effect of ovarian intra-arterial infusion of GE7-delivery system-mediated HSVl-tk/ganciclovir gene therapy in a rat model of malignant ovarian tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the gene expression of herpes simplex virus type 1 thymidine kinase (HSVl-tk) in rat malignant ovarian tumor tissues and the therapeutic effect of ganciclovior (GCV) after intra-arterial infusion of HSVl-tk gene therapy mediated by GE7-delivery system.</p><p><b>METHODS</b>A GE7-polylysine/pCMV-HSV1-tk/polylysine-HA20 4-element complex was constructed. Eighteen rats with DMBA-induced ovarian tumor were divided into 3 groups as Atk, ANS and Vtk groups. The 4-element complex GE7-polylysine/pCMV-HSV1-tk/polylysine-HA20 was injected via the ovarian artery into the rats of Atk group, saline buffer was injected in the ANS groups, and the 4-element complex was injected via the tail vein into the rats of Vtk group. All rats received intraperitoneal injection of GCV in a dose of 50 mg/kg daily for 10 days. The rats were sacrificed 3 days after the final dose of GCV, and the tumor weight was measured and tumor growth inhibition rate was calculated. Flow cytometry was used to assess the cell cycle and apoptosis.</p><p><b>RESULTS</b>The tumor weight in the rats of Atk group was (4.77 ± 2.31) g, significantly lower than that of ANS group [(14.66 ± 6.26) g, P < 0.01] and Vtk group [(17.53 ± 7.19) g, P < 0.01]. The tumor growth inhibition rate of the Atk group was 67.5%, while that of Vtk group was -19.6%. The flow cytometry showed that S-phase tumor cells in the Atk group were (54.32 ± 9.65)%, significantly higher than that in the ANS (27.43 ± 9.22)% and (30.16 ± 11.57)% in the Vtk group (both P < 0.01). The tumor cell apoptosis rate in the Atk group was (39.15 ± 12.16)%, significantly higher than that in the ANS group [(11.86 ± 5.28)%, P < 0.01] and Vtk group [(14.32 ± 6.43)%, P < 0.01].</p><p><b>CONCLUSION</b>HSV1-tk/GCV gene therapy system mediated by GE7 non-viral delivery system via ovarian arterial infusion effectively causes cell cycle arrest at S phase and enhances cell apoptosis, therefore, exerts an inhibitory effect on tumor growth.</p>

Correlation of polymorphism rs1563828 in MDM4 gene with breast cancer risk and onset age / 中华外科杂志

<p><b>OBJECTIVE</b>To explore effect of polymorphism rs1563828 (C > T) in human murine double minute 4 gene (MDM4) on genetic susceptibility for early-onset breast cancer and potential association with age of onset of breast cancer.</p><p><b>METHODS</b>One hundred and twenty-four early-onset breast cancer patients (age ≤ 35 years at time of diagnosis) from independent families admitted from January 2006 to June 2010 and 101 age-matched healthy control subjects were analyzed. Genotype analysis was conducted by polymerase chain reaction and then MALDI-TOF-MS assay. Association of genotype distribution and breast cancer risk was evaluated by χ(2) test. The odd-ratios (OR) and 95% confidence intervals (CI) were calculated by unconditional Logistic regression model. The t test was used to compare the age and demographic differences among groups.</p><p><b>RESULTS</b>The frequency of rs1563828 polymorphism genotypes in control group were CC 43.6% (44/101), CT 42.6% (43/101), TT 13.9% (14/101), and in case group were 42.7% (53/124), 46.0% (57/124), 11.3% (14/124), respectively. No significant difference (χ(2) = 0.449, P = 0.799) was reached by χ(2) test. rs1563828CT or TT genotype does not confer a significantly increased risk for breast cancer compared with CC genotype after adjusting for age, menarche in Logistic regression analysis (OR = 1.024, 95%CI: 0.581 - 1.806, P = 0.934). TT carriers were observed to develop breast cancer earlier than CC/CT carriers [(30 ± 4) years vs. (32 ± 3) years, P = 0.028].</p><p><b>CONCLUSIONS</b>The rs1563828(C > T) polymorphism in MDM4 gene may not confer risk to breast cancer, especially for early-onset breast cancer patients. Homozygous TT of rs1563828 is associated with younger age to develop breast cancer.</p>

Correlation of miR-195 with invasiveness and prognosis of breast cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the potential role of miR-195 on invasiveness and prognosis of breast cancer.</p><p><b>METHODS</b>The RNA in formalin-fixed paraffin embedded (FFPE) of 88 breast cancer patients with primary tumors was extracted, and miR-195 levels were quantified by quantitative real-time polymerase chain reaction (real-time PCR). The relationship of miR-195 levels and clinicopathological variables were assessed by Mann Whitney-U test. Recurrence-free survival and overall survival curves were derived from Kaplan-Meier estimates and the curves were compared by Log-rank tests. Cox regression analysis was used for multivariate analysis. All statistical tests were two-sided.</p><p><b>RESULTS</b>The levels of miR-195 in the breast cancer with histological high grade, tumor size of T3-4, lymph nodal involvement or vessel invasion were significantly down-regulated, compared with those of patients with histological low grade (Z = -2.271, P = 0.023), tumor size of T1-2 (Z = -2.687, P = 0.007), no lymph node metastasis (Z = -1.967, P = 0.049) and vessel invasion (Z = -2.432, P = 0.015). In addition, no statistically significant difference (P > 0.05) was identified between miR-195 levels and hormone receptors status, HER-2 expression, TNM stage, tumor types, recurrence and menstrual status. When considering 2(-ΔCt) = 0.270 (median level) as cut-off value, Kaplan-Meier survival analysis indicated that patients with high miR-195 level showed a positive association towards a longer survival, either recurrence-free survival (χ(2) = 5.985, P = 0.014) or overall survival (χ(2) = 30.05, P = 0.000). In a multivariate analysis, miR-195 expression on FFPE correlated significantly with outcomes of breast cancer (HR = 0.040, 95%CI: 0.009 - 0.179, P = 0.000) and was independent of other prognostic factors.</p><p><b>CONCLUSIONS</b>It suggests that miR-195 expression on FFPE is inversely correlated with histological high grade, bigger tumor size, lymph node involvement, vessel invasion. Furthermore, as independent prognostic factor, low miR-195 significantly contributes to poor outcomes of breast cancer.</p>

Correlation of miR-155 on formalin-fixed paraffin embedded tissues with invasiveness and prognosis of breast cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the potential use of miR-155 as novel breast cancer biomarker.</p><p><b>METHODS</b>There were 88 breast cancer patients underwent modified mastectomy and had detailed clinical follow-up information. Extracting RNA from the formalin-fixed paraffin embedded (FFPE) samples, miR-155 levels were quantified by real-time-PCR. miR-155 levels among clinico-pathological variables were accessed by Mann Whitney-U test. Overall survival curve was derived from Kaplan-Meier estimates and the curve was compared by Log-rank test. Cox regression analysis was used for multivariate analysis. All statistical tests were two-sided.</p><p><b>RESULTS</b>Significantly higher miR-155 level was found in tumor tissue compared to paired normal tissue (t = 6.75, P = 0.000). A potential relationship between miR-155 levels and existing clinico-pathological parameters of breast cancer, such as menstrual status, tumor size, nodal involvement, stage of disease, hormone receptor status, HER-2 status, histological grade or tumor subtype was investigated. Up-regulated miR-155 level was observed in breast cancer with lymph node metastasis, pT3+4, advanced TNM stage, HER-2 positive and with vascular invasion (Z = -6.320 to -2.041, P = 0.000 to 0.041). When considering 2(-ΔCt) = 4.87 (median level) as cut-off value, patients with miR-155 up-regulation showed a positive association towards a shorter overall survival (χ(2) = 6.396, P = 0.011). In Cox multivariate analysis, miR-155 expression on FFPE was shown an inverse trend for outcomes of breast cancer (HR = 1.58, 95%CI: 0.87 - 3.16, P = 0.082).</p><p><b>CONCLUSIONS</b>miR-155, as an oncomir, promotes lymph node involvement and vascular invasion and accompanies over-expressed HER-2 on breast cancer FFPE tissue. It suggests that miR-155 could predict the invasiveness.</p>

Lapatinib plus capecitabine in treating HER2-positive advanced breast cancer: efficacy, safety, and biomarker results from Chinese patients / 癌症

Overexpression of human epidermal growth factor receptor-2 (HER2) in metastatic breast cancer (MBC) is associated with poor prognosis. This single-arm open-label trial (EGF109491; NCT00508274) was designed to confirm the efficacy and safety of lapatinib in combination with capecitabine in 52 heavily pretreated Chinese patients with HER2-positive MBC. The primary endpoint was clinical benefit rate (CBR). Secondary endpoints included progression-free survival (PFS), time to response (TTR), duration of response (DoR), central nervous system (CNS) as first site of relapse, and safety. The results showed that there were 23 patients with partial responses and 7 patients with stable disease, resulting in a CBR of 57.7%. The median PFS was 6.34 months (95% confidence interval, 4.93-9.82 months). The median TTR and DoR were 4.07 months (range, 0.03-14.78 months) and 6.93 months (range, 1.45-9.72 months), respectively. Thirteen (25.0%) patients had new lesions as disease progression. Among them, 2 (3.8%) patients had CNS disease reported as the first relapse. The most common toxicities were palmar-plantar erythrodysesthesia (59.6%), diarrhea (48.1%), rash (48.1%), hyperbilirubinemia (34.6%), and fatigue (30.8%). Exploratory analyses of oncogenic mutations of PIK3CA suggested that of 38 patients providing a tumor sample, baseline PIK3CA mutation status was not associated with CBR (P = 0.639) or PFS (P = 0.989). These data confirm that the lapatinib plus capecitabine combination is an effective and well-tolerated treatment option for Chinese women with heavily pretreated MBC, irrespective of PIK3CA status.

Evaluation of GE7-transferring system-mediated HSV(1)-tk gene transfer in a rat model of ovarian tumor via intra-arterial route / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the gene and protein expression of herpes simplex virus type I-thymidine kinase (HSV(1)-tk) in the ovarian tumor tissues and other organs after arterial infusion of HSV(1)-tk gene mediated by GE7 delivery system.</p><p><b>METHODS</b>GE7-polylysine/pCMV-HSV(1)-tk/polylysine-HA20 complexes were constructed. Nine rats with induced ovarian tumor were divided into 3 groups, injecting the 4-element complexes or saline buffer through the ovarian artery and complexes through the tail vein, respectively. The ovarian tumors, hearts, livers, spleens, lungs and kidneys were obtained at 72 hours after injection. RT-PCR and Western Blot were preceeded to determine the expression of HSV(1)-tk gene and protein in the tumor tissues and other organs.</p><p><b>RESULTS</b>In the group of arterial injection with 4-element complexes, the HSV(1)-tk gene and protein were expressed strongly in the tumor tissues, while little or none was detected in other organs. In the group of arterial injection with saline buffer, no HSV(1)-tk gene and protein was detected in both tumor tissues and other organs. In the group of tail vein injection, none was detected in tumor tissues and only little was found in the livers, spleens, lungs and kidneys.</p><p><b>CONCLUSION</b>High target and gene transfer rates can be obtained when HSV(1)-tk gene is transferred via the artery route mediated by GE7 delivery system. HSV(1)-tk protein can be expressed after the gene transfer. The results may provide a new strategy for target killing of HSV(1)-tk/GCV system in ovarian tumors.</p>

A prospective comparison of molecular assay and touch imprint cytology for intraoperative evaluation of sentinel lymph nodes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Accurate intraoperative diagnosis of sentinel lymph node (SLN) metastases enables the selection of patients for axillary lymph node dissections during the same operation, reducing the need for a second operation. The present study aimed to prospectively compare the GeneSearch(TM) Breast Lymph Node (BLN) Assay with touch imprint cytology (TIC) for intraoperative evaluation of SLNs.</p><p><b>METHODS</b>SLNs were sectioned in 1.5 - 3.0 mm pieces. TIC was performed on all pieces and the BLN Assay and postoperative histology evaluations were performed on different alternating node pieces. Overall performance of the BLN Assay was compared with that of TIC relative to the postoperative histology results.</p><p><b>RESULTS</b>A total of 90 patients enrolled in the study. Complete intraoperative data for both the BLN Assay and TIC were collected in 86 patients. The sensitivity, specificity, and overall accuracy of the BLN Assay were 82%, 97%, and 92%, respectively on a per patient basis compared with those of TIC which were 67%, 100%, and 90%.</p><p><b>CONCLUSIONS</b>Performance of the BLN Assay was superior to that of TIC and the additional application of TIC did not help improve the total sensitivity and accuracy of the intraoperative assessment. The existence of ectopic breast tissue might be a possible cause of false positive for the BLN assay. In addition, the BLN Assay complements histopathology assessment and can minimize sampling error without increasing pathologists' workload.</p>

Risk factors for postoperative seromas in Chinese breast cancer patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Seroma formation is one of the most common complications after breast cancer surgery. Various risk factors have been evaluated for their associations with the development of seromas in Western populations. However, similar data are not available in Chinese series. Therefore, we sought to investigate the potential risk factors for Chinese breast cancer patients.</p><p><b>METHODS</b>A prospective study of female breast cancer patients undergoing surgery was carried out in Cancer Hospital of Fudan University, Shanghai, China. Univariate analyses were performed by chi-square test or Student's t test or Mann-Whitney test and multivariate analyses by stepwise Logistic regression. The logistic model included age (years), total serum protein concentration (g/L), drainage volume on postoperative day 3 (POD 3; ml) and time to daily drainage volume not more than 30 ml (TTV30; days).</p><p><b>RESULTS</b>A total of 158 patients with breast cancer were studied. The mean age at diagnosis was (52.14 ± 10.77) years (range 25 - 92). During the follow-up period, 24 (15.2%) patients developed seromas. Calculated as continuous variables in the stepwise Logistic regression, age (OR = 1.090, 95%CI 1.028 - 1.155, P = 0.004), total serum protein concentration (OR = 0.886, 95%CI 0.791 - 0.992, P = 0.036), drainage volume on POD3 (OR = 1.013, 95%CI 1.002 - 1.023, P = 0.017) and TTV30 (OR = 1.273, 95%CI 1.039 - 1.561, P = 0.020) were independent risk factors for seroma formation. Additionally, significant difference in daily drainage volume was substantiated in the analysis by seroma formation (P = 0.034) rather than by type of surgery (P = 0.713).</p><p><b>CONCLUSIONS</b>Although the pathogenesis of seroma remains controversial, such risk factors as age, nutritional status, drainage volume on POD3 and TTV30 should be considered for prediction and prevention of seroma formation in Chinese breast cancer patients.</p>

Retrospective analysis of trastuzumab treatment in 141 patients with Her-2 positive breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To summarize the clinical experience of trastuzumab treatment in neoadjuvant, adjuvant, metastatic setting of Chinese patients with Her-2 positive breast cancer and evaluate the efficacy of trastuzumab in combination with chemotherapy.</p><p><b>METHODS</b>From January 2004 to December 2008, 141 outpatients with breast cancer treated with trastuzumab were investigated retrospectively. The follow-up time ranged from 3 to 319 months. The disease free survival time (DFS) of metastatic setting was calculated. The overall survival time (OS), time to treatment failure (TTF) and clinical response rate (CRR, including complete response, partial response and stable disease) of adjuvant, first-line, second-line therapy were analyzed statistically.</p><p><b>RESULTS</b>In the neoadjuvant regimen, paclitaxel plus carboplatin in combination with trastuzumab accounted for 66.7%, which achieved pathological complete response in 10 of 16 patients. In the adjuvant regimen, anthracycline or anthracycline followed by taxane accounted for 53.9%. The median DFS of 57 cases with metastatic diseases was 17 months. The CRR of first-line trastuzumab use in metastatic setting was 84.5%, compared with 44.4% of second-line use. The median TTF of first-line treatment was 24 months compared with 5 months of second-line treatment. Statistically significant differences were observed.</p><p><b>CONCLUSION</b>The regimen of paclitaxel plus carboplatin in combination with trastuzumab deserves wide clinical use. In metastatic setting, first-line treatment of trastuzumab plus chemotherapy can achieve a higher response rate than second-line treatment. Continued trastuzumab therapy combined with different chemotherapy treatment after disease progression may obtain additive clinical advantage.</p>

Analysis of BRCA1 gene mutations in patients with early-onset breast cancer and their affected relatives in Guangdong province / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the BRCA1 mutations in patients with early-onset breast cancer and their affected relatives in Guangdong province and explore the relationship between BRCA1 mutation and the expressions of estrogen receptor(ER), progesterone receptor(PR), HER2 and ALN.</p><p><b>METHODS</b>From 58 patients with early-onset breast cancer and their affected relatives, the genomic DNA was extracted from the peripheral blood mononuclear cells and the coding regions of the BRCA1 gene was amplified using polymerase chain reaction. BRCA1 gene mutations were screened by denaturing high performance liquid chromatography (DHPLC) and subsequent direct DNA sequencing. The expression of ER, PR, HER2 and ALN were detected with immunohistochemistry and their relations with the gene mutation were analyzed.</p><p><b>RESULTS</b>Disease-related BRCA1 mutations were detected in 2 of the 58 patients, who were younger than 35 years old, including 1 with a novel splice-site mutation (IVS5-1 G-->A). No association was found between this novel mutation and the expressions of ER, PR, HER2 and ALN.</p><p><b>CONCLUSION</b>The incidence of BRCA1 mutation is significantly lower in patients with early-onset breast cancer and their affected relatives in Guangdong province than in the Western populations. The novel mutation identified in BRCA1 gene may represent a mutation characteristic of the patients in Guangdong province. BRCA1 gene mutations may not have any relation with the expression of ER, PR, HER2 and ALN.</p>

Study on predictors of long term results for neo-adjuvant chemotherapy in locally advanced breast cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To identify predictive markers of the long-term outcome for neo-adjuvant chemotherapy (NC) in locally advanced breast cancer (LABC) treated with intravenous vinorelbine (V) and epirubicin (E) combination regimen.</p><p><b>METHODS</b>One hundred and nineteen patients with LABC were treated from September 2001 to May 2006. All patients were diagnosed as invasive breast cancer by 14G core needle biopsy and treated with three cycles of VE regimen before the operation. The patients were subjected to surgery and subsequently were given other three cycles of VE or cyclophosphamide+epirubicin+fluorouracil (CEF) regimen according to the clinical responses. Local-regional radiotherapy was applied to all patients after the chemotherapy and followed by hormone-therapy according to hormone receptor status. The impact of clinical, pathological, and immunohistochemical features on disease free survival (DFS) and overall survival (OS) was evaluated.</p><p><b>RESULTS</b>All patients were evaluable for responses: clinical complete response was documented in 27 patients (22.7%), 78 patients (65.5%) obtained partial clinical response. The pathological complete response was found in 22 cases (18.5%). Of the patients, 115 cases (96.6%) were followed-up for a median time of 63.4 months (range, 9-76 months), the 5-year DFS rate and OS rate was 58.7% and 71.3%, respectively. On multivariate analysis, high pre-Ki-67 (P=0.012) and post-Ki-67 expression (P=0.045), no pathological complete response after NC (P=0.034) were associated with the higher risk of disease relapse; high pre-Ki-67 (P=0.017) and post-Ki-67 expression (P=0.001), negative pre-ER (P=0.002) and no pathological complete response after NC (P=0.034) were associated with a shorter survival.</p><p><b>CONCLUSION</b>Pathological response in primary tumor, pre-Ki-67 and post-Ki-67 expression, pre-ER expression are important predictors of long-term outcome for LABC patients with three cycles of VE regimen before operation.</p>

A retrospective study of 129 cases with immediate breast reconstruction after skin-sparing mastectomy for breast cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the oncologic safety, indications and aesthetic results for skin-sparing mastectomy (SSM) and immediate breast reconstruction (IBR).</p><p><b>METHOD</b>One hundred and twenty-nine breast cancer patients treated by SSM + IBR from October 1999 to May 2007 were reviewed. Reconstructive techniques included latissimus dorsi flaps (38 patients), implants only (2 patients), latissimus dorsi flaps plus implants (61 patients), pedicled transverse rectus abdominis myocutaneous (TRAM) flaps (25 patients) and deep inferior epigastric artery perforator (DIEP) flaps (3 patients). Aesthetic results were judged by patients' self-evaluation.</p><p><b>RESULTS</b>Mean duration of hospitalization was 18.6 days. Time of first chemotherapy was 5.2 days after operation. Eleven patients (11/63, 17.5%) developed capsular contracture and 24 patients (24/99, 24.2%) developed seroma in the donor site. Nine patients (9/28, 32.1%) developed partial fat necrosis in TRAM and DIEP flaps. The satisfaction with the aesthetic results of the reconstructive breast was significantly lower in irradiated patients than non-irradiated ones. Median follow-up time was 11 months. Five patients developed local recurrence and 7 patients with metastasis.</p><p><b>CONCLUSIONS</b>SSM with IBR can be used for the 0 to II a stage breast cancer patients, with surgical oncologic and aesthetic satisfaction. Radiotherapy has an adverse effect on the reconstructive breast. Delayed or delayed-immediate reconstructions are recommended for patients indicated to postoperative radiotherapy.</p>

Expression of CK5/6 and CK17 and its correlation with prognosis of triple-negative breast cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the correlation of CK5/6 and CK17 expression with clinical outcome in patients with triple-negative [ER(-), PR(-), Her-2(-)] breast cancer.</p><p><b>METHODS</b>112 patients with breast cancer treated by surgery between 2000 and 2002 were included in this study. All cases were immunohistochemically proven to be triple-negative. Samples of formalin-fixed and paraffin-embedded surgical specimens were obtained for immunohistological examination for CK5/6 and CK17 expression. The correlation of the gene expression with clinicopathological features and outcome of the patients was analyzed.</p><p><b>RESULTS</b>Of the 112 triple-negative patients, five-year disease-free survival rate was 73.2% (82/112). The positive rate of both CK5/6 and CK17 was 21.4% (24/112), either CK5/6 or CK17 positive was 46.4% (52/112). It was shown by Kaplan-Meier curve that positive CK5/6, CK17 or CKs (CK5/6 or CK17 positive) was correlated with poor five-year disease-free survival (P = 0.020, P = 0.032, P = 0.003); and positive staining of CK5/6 or CKs was correlated with poor five-year overall survival (P = 0.027, P = 0.015). Of the 91 patients with invasive ductal carcinoma, a correlation of CK5/6 or CK17 positive staining with high grade differentiation was observed (P = 0.030), and with axillary lymph node metastasis was also noticed (P = 0.044). Multivariate analysis by Cox regression showed that differentiation grade, pathological stage and expression of CK5/6 were factors affecting both the disease-free-survival and overall-survival, while menopausal status was an independent factor affecting the disease-free-survival.</p><p><b>CONCLUSION</b>Positive expression of CK5/6 or CK17 in patients with triple-negative breast cancer is correlated with poor prognosis, high grade differentiation and axillary lymph node metastasis.</p>

Surgical management of early breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the available surgical treatment modalities so as to explore the optimal strategy of managing early breast cancer.</p><p><b>METHODS</b>The clinical data of 2173 consecutive early-stage breast cancer patients treated by surgery treatments were retrospectively reviewed in order to clarify the indications and contraindications of different modalities. Therapeutic outcome of different surgical treatment modes were compared in terms of recurrence-free survival ( RFS) , disease-free survival ( DFS) , overall survival (OS). The cosmetic results of breast conservation and reconstruction were also evaluated .</p><p><b>RESULTS</b>The median age of these patients was 51 years ranging from 18 to 91. Of 2173 patients, 547 had stage 0- I lesions and 1626 stage II , and 1155 (53. 2% ) premenopausal. The proportion of patients who received radical surgery, breast conservation and reconstruction after mastectomy was 83. 6% (1817/2173), 10. 5% (229/2173) and 2. 5% (55/2173) , respectively. Younger and premenopausal patients prefer conservative and reconstructive surgeries, which are reasonable for stage 0-I and non-invasive breast cancer patients. Conservative surgery was not suitable for Paget's disease of breast (P = 0. 004) , mastectomy followed by reconstruction in this type of cancer was up to 38. 5%. The recurrence and metastasis rate of conservation or mastectomy were similar with a comparable 3-year RFS of 97. 4% and 95. 4% , respectively; there were also no significant differences in RFS(P =0. 2435) , DFS( P =0. 1395) and OS(P =0. 9406) after having been followed for 3 to 64 months. Similarly, immediate reconstruction did not show any negative effects with only 1 recurrence and 1 metastasis. Aesthetic outcomes were assessed as excellent or good in 90. 0% of breast conservation surgery, and the acceptability of reconstruction was 94. 5%.</p><p><b>CONCLUSION</b>Breast conserving surgery not only has comparable survival as mastectomy, but also has better cosmetic outcomes. Immediate breast reconstruction can be a suitable option without compromising survival. It is very important in the management for early breast cancer by selecting the most suitable surgery mode for every individual patient not only to cure her disease but also to satisfy the patient psychologically. Conservation should be preferred prior to reconstruction whenever possible.</p>

Evaluation of intraoperative touch imprint cytology of sentinel lymph node for breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the intraoperative touch imprint cytology as an diagnostic method of sentinel lymph node for breast cancer patient.</p><p><b>METHODS</b>Sentinel lymph node biopsy was performed in 105 selected early breast cancer patients, and sentinel lymph node was identified in 101 (96.19%) of these patients. Axillary lymph node dissection was also performed in almost all the patients. All the sentinel lymph nodes were cut into 2-3 mm pieces along the long axis. Touch imprint was made of each piece of the sentinel lymph node, then air-dried, and finally stained with H&E. Intraoperative touch imprint cytology results were compared with the final paraffin H&E pathology. All sentinel nodes were cut into 4 microm sections every 100-microm interval, and the series sections were stained with H&E.</p><p><b>RESULTS</b>202 sentinel lymph nodes were identified in 101 breast cancer patients. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value of intraoperative imprint cytology for 202 sentinel nodes was 92.1%, 98.8%, 97.5%, 94.6% and 98.2%, respectively; which was 89.3%, 98.6%, 96.0%, 96.2% and 96.0%, respectively in the 101 patients with identified sentinel node. Compared with the series sections, the sensitivity, specificity, accuracy, positive predictive value, negative predictive value of intraoperative imprint cytology for sentinel nodes was 83.3%, 98.8%, 95.5%, 94.6% and 95.8%, respectively; and it was 81.3%, 100.0%, 94.1%, 100.0% and 92.0%, respectively in 101 patients with identified sentinel node.</p><p><b>CONCLUSION</b>Touch imprint cytology is a simple, effective and rapid method for intraoperative pathological evaluation of sentinel lymph node for breast cancer patient, which has a high concordance with the paraffin results, and can provide accurate and rapid diagnosis information for the surgeon during operation.</p>

5589del8: the recurrent mutation of BRCA1 gene in Chinese breast cancer patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the "hot spot" of BRCA1/2 gene mutations in Chinese mainland breast cancer population.</p><p><b>METHODS</b>The known BRCA1/2 gene mutations in author's previous studies were reanalyzed by denaturing high performance liquid chromatography and DNA sequencing method in 177 patients with early onset breast cancer or affected relatives and 426 sporadic breast cancer patients from four breast cancer centers in China.</p><p><b>RESULTS</b>Three cases were found with BRCA1 5589del8 mutation out of 247 hereditary-predisposing breast cancer patients (70 patients in previous study and 177 patients in current study) and 2 cases with BRCA1 5589del8 mutation out of 426 sporadic breast cancer patients. They had similar even same haplotype.</p><p><b>CONCLUSION</b>BRCA1 5589del8 mutation is likely to be the "founder mutation" in Chinese population, but it should be confirmed by further studies.</p>

BRCA1 germ line mutations in Chinese early-onset breast cancer patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the role of disease associated germ line mutations in BRCA1 gene among Chinese early-onset breast cancer patients.</p><p><b>METHODS</b>A total of 188 early-onset breast cancer patients, who were diagnosed with breast cancer before 41-year-old, were enrolled from four breast cancer clinical centers in China. Thirty-nine of them (20.7%) also had family history of breast/ovarian cancer. DNA extracted from lymphocytes was amplified by polymerase chain reaction (PCR) for the entire exons and the splicing sites of BRCA1. Twenty-two of the patients were screened by single strand conformation polymorphism (SSCP), and the other 166 of them were screened by denaturing high performance liquid chromatography (DHPLC). The abnormal fragments recognized were ascertained by DNA direct sequencing. For those samples with the same recurrent mutation, five BRCA1-linked markers (D17S855, D17S1322, D17S1323, D17S1326 and D17S1327) were used for the allelotype analysis.</p><p><b>RESULTS</b>Twelve disease-associated mutations were identified in 15 (8.0%) patients, among which BRCA1 1100delAT and 5589del8 were identified in 3 and 2 patients respectively. Nine (23.1%) of them were identified in those with breast/ovarian cancer family history. The difference of BRCA1 mutation frequency between the patients with and without family history was statistically significant (P=0.001). Allelotype analysis showed the two BRCA1 5589del8 mutation carriers shared the same allelotype in all the 5 STR sites, and two of the three 1100delAT mutation carriers, who came from the northern China, also shared the same allelotype in all the 5 STR sites, which were different from those of the 5589del8 mutation carriers'.</p><p><b>CONCLUSION</b>This is a relatively very large scale multi-hospital-based study of BRCA1 mutations in Chinese early-onset breast cancer patients up to now. It seems reasonable to give genetic consultations and genetic test of BRCA1 gene to early-onset breast cancer patients in China, especially for those with breast/ovarian cancer family history. The two recurrent mutations might be founder mutations of Chinese population. It might be cost-effective to analyze these two mutations before whole gene analysis.</p>

Suicidal cancer vaccine enhances anti-tumor immunotherapeutic effect and its safety in the treatment of ovarian cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the anti-tumor immunotherapeutic effect induced by the suicidalcancer vaccine FC/TK, and to evaluate the safety of this vaccine.</p><p><b>METHODS</b>The suicidal cancer vaccine, named FC/TK, was prepared by fusion of suicide gene (HSVI,-TK gene) -modified ovarian carcinoma NuTu-19 cells with rat bone marrow-derived dendritic cells (DCs). The morphology of FC/TK was evaluated by scanning electron microscopy. The stimulatory effect of FC/TK on T cells was determined by T cell proliferation assay. In immunotherapeutic studies in vivo, Fischer344 rats were injected subcutaneously with NuTu-19 cells, followed by treatment of FC/TK on days 7 and 14, compared to controls treated with irradiated FC/TK, FC or PBS, respectively. Tumor incidence and volume were measured in 90 days after challenge. To determine the killing effect of FC/TK in vivo, TUNEL assays were applied to detect apoptotic cell death in spleen of vaccinated rats with prodrug ganciclovir administration.</p><p><b>RESULTS</b>FC/TK cells were of irregular shape with surface membrane processes. Compared to the control groups, FC/TK significantly promoted T cell proliferation (P <0.01). The rats vaccinated with FC/TK and FC significantly inhibited the tumor growth compared to rats vaccinated with irradiated FC/TK (P <0.05) or with PBS ( P <0.01). The immunotherapeutic effect induced by FC/TK was similar to that using FC. Fluorescence microscopy showed that fluorescein-stained FC/TK cells migrated into spleen also showed to be TUNEL-positive, suggesting that the FC/TK cells were killed by ganciclovir in vivo.</p><p><b>CONCLUSION</b>Our data indicate that suicidal cancer vaccine is an effective and safe therapy for ovarian carcinoma and may serve as a broadly applicable approach for other cancer vaccines in the future.</p>

Expression of ER alpha in chemically induced MDA-MB-435 cells and its responsiveness to endocrine / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of ER alpha in chemically induced, ER alpha-negative human breast cancer MDA-MB-435 cells and its restoration of the responsiveness to endocrine therapy.</p><p><b>METHODS</b>MDA-MB-435 cells were treated with HDAC inhibitor trichostatin A(TSA)and DNMT1 inhibitor 5-AZA-CdR (AZA). The mRNA level of ER alpha, PR and PS2 in treated MDA-MB-435 cells was detected by RT-PCR. The WST-8 (water-soluble tetrazolium salt-8) method was used to analyze the proliferation rate of the cells. Xenograft in female nude mice was used to further explore the change of proliferation rate of treated MDA-MB-435 cells in vivo.</p><p><b>RESULTS</b>After treatment with AZA and TSA, mRNA expression of ER alpha, PR and pS2 was up-regulated in MDA-MB-435 cells. The mRNA level of ER alpha was the hightest when MDA-MB-435 cells were treated with 2.5 micromol/L AZA and 100 ng/ml TSA. The treated MDA-MB-435 cells showed different proliferation rate in various media containing different concentration of estrodial. The MDA-MB-435 cells showed down-regulated proliferation rate after treatment with the combination of 2.5 micromol/L AZA and 100 ng/ml TSA, and 4-OH tamoxifen could suppress the growth rate of the induced MD-MBA-435 cells but not the untreated cells. The treated MDA-MB-435 cells showed slower proliferation rate than that of untreated cells in vivo (P <0. 01), and the proliferation rate of the treated MDA-MB-435 cells became lower when the nude mice were deprived of estrogen by castration (P <0. 01).</p><p><b>CONCLUSION</b>After treatment with TSA and AZA, ER alpha-negative MDA-MB-435 cells can express functional ER alpha and regain responsiveness to estrogen both in vitro and in vivo. HDAC inhibitor and DNMT1 inhibitor may play an important role in restoration of sensitivity of ER alpha-negative breast cancers to endocrine therapy.</p>

BRCA1 and BRCA2 gene mutations of familial breast cancer from Shanghai in China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of BRCA1 and BRCA2 gene mutations among breast cancer patients with affected relatives in Shanghai of China.</p><p><b>METHODS</b>Thirty-five breast cancer patients who had at least one first-degree relative affected were analyzed, among whom 13 patients suffered from breast cancer at age of less than 40 years. A comprehensive BRCA1 and BRCA2 mutation analysis was performed through denaturing high-performance liquid chromatography (DHPLC) and subsequent DNA direct sequencing.</p><p><b>RESULTS</b>Four mutations in BRCA1 gene, including 2 novel splice-site mutations (IVS17-1G>T, IVS21+1G>C) and 2 frameshift mutations (1100delAT; 5640delA) were identified. One frameshift mutation (5802delAATT) was detected in exon 11 of BRCA2. Additional 12 novel single nucleotide polymorphisms(SNPs) were detected, including a novel unclassified variant and 7 novel intronic variants in BRCA1, and 4 novel intronic variants in BRCA2, with which all caused no alteration of amino acid coding. The mutation frequency of BRCA1 and BRCA2 in patients with family history was 11.4% and 2.9%, respectively.</p><p><b>CONCLUSION</b>Two novel mutations in BRCA1 may be mutations characterized to familial breast cancer of Chinese Shanghai population. The BRCA2 may contribute to mutation less than BRCA1 in familial breast cancer. Our data contribute to information on mutation spectrum of BRCA gene in Chinese population and also offer a recommended screening mode for clinical genetic testing policy in China.</p>