Long-Term Outcomes of Orthotopic Neobladder Versus Ileal Conduit Urinary Diversion in Robot-Assisted Radical Cystectomy (RARC): Multicenter Results from the Asian RARC Consortium.

PURPOSE: Robot-assisted radical cystectomy (RARC) has gained traction in the management of muscle invasive bladder cancer. Urinary diversion for RARC was achieved with orthotopic neobladder and ileal conduit. Evidence on the optimal method of urinary diversion was limited. Long-term outcomes were not reported before. This study was designed to compare the perioperative and oncological outcomes of ileal conduit versus orthotopic neobladder cases of nonmetastatic bladder cancer treated with RARC. PATIENTS AND METHODS: The Asian RARC consortium was a multicenter registry involving nine Asian centers. Consecutive patients receiving RARC were included. Cases were divided into the ileal conduit and neobladder groups. Background characteristics, operative details, perioperative outcomes, recurrence information, and survival outcomes were reviewed and compared. Primary outcomes include disease-free and overall survival. Secondary outcomes were perioperative results. Multivariate regression analyses were performed. RESULTS: From 2007 to 2020, 521 patients who underwent radical cystectomy were analyzed. Overall, 314 (60.3%) had ileal conduit and 207 (39.7%) had neobladder. The use of neobladder was found to be protective in terms of disease-free survival [Hazard ratio (HR) = 0.870, p = 0.037] and overall survival (HR = 0.670, p = 0.044) compared with ileal conduit. The difference became statistically nonsignificant after being adjusted in multivariate cox-regression analysis. Moreover, neobladder reconstruction was not associated with increased blood loss, nor additional risk of major complications. CONCLUSIONS: Orthotopic neobladder urinary diversion is not inferior to ileal conduit in terms of perioperative safety profile and long-term oncological outcomes. Further prospective studies are warranted for further investigation.

Intravesical hyaluronidase causes chronic cystitis in a rat model: a potential model of bladder pain syndrome/interstitial cystitis.

OBJECTIVES: To determine whether a potential rat model of bladder pain syndrome could be developed through long-term intermittent intravesical hyaluronidase. METHODS: A total of 64 female Sprague-Dawley rats were divided into a control group, a low-dose hyaluronidase (1 mg/mL) group, a high-dose hyaluronidase (4 mg/mL) group and a hyaluronic acid-treated group. Hyaluronidase was given intravesically three times a week for 1 month. Hyaluronic acid (0.5 mL, 0.8 mg/mL) was introduced intravesically to hyaluronidase-treated rats' bladders. Histological changes, cystometry, nociceptive behaviors, and messenger ribonucleic acid levels of inflammatory factors were evaluated and compared between groups. RESULTS: All hyaluronidase-treated rats showed chronic inflammation and fibrosis, increased and activated mast cells, thinned bladder epithelium with abnormal expressions of uroplakin III and zonula occluden-1, and increased levels of interleukin-6 and intercellular adhesion molecule-1 messenger ribonucleic acid. However, the inflammatory score and levels of interleukin-6 and intercellular adhesion molecule-1 were more significant in the high-dose hyaluronidase group than in the low-dose hyaluronidase group (P < 0.01). Furthermore, hyaluronidase-treated rats showed markedly decreased intercontraction intervals, bladder capacity and increased sensitivity to pain compared with controls (P < 0.01). Hyaluronic acid treatment significantly decreased the inflammatory level, number of mast cells, sensitivity to pain, levels of interleukin-6 and intercellular adhesion molecule-1, and increased intercontraction intervals and bladder capacity (P < 0.01). CONCLUSIONS: Long-term intermittent intravesical hyaluronidase could develop a severe chronic cystitis with diffused fibrosis accompanied by altered histology and bladder function. This chronic cystitis rat model can resemble the clinical and histopathological features of human bladder pain syndrome, and might be a potential valuable model for investigation of this troublesome disease.

Percutaneous nephrolithotomy in semisupine position: a modified approach for renal calculus.

Conventional percutaneous nephrolithotomy (PCNL) is usually performed in a prone position, which compresses the thorax and results in difficulty in rescue during operation. When PCNL is performed in a supine position, the flank renal puncture area is limited, so it is difficult to treat disseminated and complex renal calculi. Herein, we introduce a modified semisupine position for performing PCNL, which has numerous benefits as well as safe and effective. Between May 2002 and May 2009, a total of 452 patients with renal calculi were treated with semisupine PCNL. The patient was placed in 45° semisupine position during the procedure, with the affected flank arched as much as possible. In this series, no one converted to open surgery. The average operating time was (115.2 ± 44.5) min. Single tract PCNL was performed for 80.97% of the cases, two tracts 13.94%, three tracts 4.65%, and four tracts 0.44%. The upper, middle, and lower calix tracts accounted for 12.1, 63.0, and 24.9%, of procedures, respectively. Stone-free rate was 85.7% overall, 92.2% for single calculus (83/90), and 72.9% for staghorn calculi (78/107). Major postoperative complications occurred in 3.3% of the cases. This study demonstrated PCNL in a semisupine position is an effective alternative for treating renal calculi, which combines the advantages of PCNL in a prone position, and PCNL in a supine position. The semisupine position allows easier irrigation of stone fragments, is more comfortable for the patient, and facilitates monitoring of anesthesia.

[Inhibition and significance of pigment epithelium-derived factor in the development and metastasis of prostate cancer].

OBJECTIVE: To study the inhibition and significance of pigment epithelium-derived factor (PEDF) in the development and metastasis of prostate cancer. METHODS: The expression of PEDF was examined in the normal prostate tissue, benign prostatic hyperplasia, prostate cancer tissue and prostate cancer cell lines, PC-3 and Lncap by immunohistochemical SP method and Western blot. In combination with clinical data, statistical analysis was performed to evaluate the relation of the expression level of PEDF in prostate cancer and the relationship between different histological grades of prostate cancer. RESULTS: In normal prostate tissue and benign prostate tissue, the expression of PEDF were elevated and it was far higher than the prostate cancer and prostate cancer cell line. The expression of PEDF and the pathological grade of prostate cancer were related to the differentiated carcinoma of prostate tissue, and the expression level of PEDF in poorly differentiated carcinoma below the highly differentiated carcinoma of prostate cancer. In metastatic prostate cancer, the expression of PEDF was lower than that of prostate cancer without metastasis (12% vs 43.1%). CONCLUSION: The expression of PEDF and the incidence of prostate cancer have a negative correlation. The lower grade of prostate cancer cells, the less the tissue expression of PEDF and the higher tendency of invasion and metastasis.

Laparoscopic radical cystectomy with orthotopic ileal neobladder in the female: report of 14 cases.

BACKGROUND: Bladder carcinoma is the most common malignant urological tumor in China. We present our preliminary experience and results of laparoscopic radical cystectomy (LRC) with orthotopic ileal neobladder in female patients with bladder carcinoma. METHODS: From February 2003 to February 2008, 14 female patients with bladder carcinoma underwent LRC with orthotopic ileal neobladder. Nine of these patients underwent hysterectomy and ovariectomy, and the other 5 had preservation of the uterus and ovarian appendage. Standard bilateral pelvic lymphadenectomy was followed by radical cystectomy that was completed laparoscopically with hysterectomy and ovariectomy when needed. The tumor was removed by a 4 - 5 cm lower midline abdominal incision, followed by the construction of ileal neobladder and the extracorporeal anastomosis of ureter-neobladder. The neobladder was anastomosed to the urethral stump under a laparoscope. RESULTS: The mean operative time and blood loss in the 14 patients were 350.2 minutes and 349.8 ml, respectively. Postoperative complications included uretero-pouch anastomotic stricture in 1 patient and pouch-vaginal fistula in 1 patient. Follow-up time of all patients ranged from 3 to 60 months, and 12 patients were followed up for more than 6 months and achieved micturition in half a year. One patient had occasional day-time urinary incontinence and 2 had night-time incontinence. Two patients who had undergone hysterectomy and ovariectomy had voiding difficulties after one year, which was treated by intermittent self-catheterization. The mean volume of the neobladder and the residual urine were 333.6 ml and 31.2 ml, respectively. Surgical margins were tumor free for all patients. One patient had bone metastasis and died 11 months after the operation. CONCLUSIONS: LRC with orthotopic ileal neobladder in female patients is a technically feasible, safe and mini-invasive procedure with a low morbidity and acceptable neobladder function. Long-term follow-up is required to confirm the neobladder function and oncological outcomes.

[Laparoscopic radical cystectomy with orthotopic ileal neobladder: report of 108 cases].

OBJECTIVE: To analyze the effects, complication, and outcome of laparoscopic radical cystectomy. METHODS: 108 patients with bladder cancer, 96 males and 12 females, aged 61 (36 - 81) underwent laparoscopic radical cystectomy with orthotopic ileal neobladder. Five-port transperitoneal approach was applied. The surgical procedure included standard laparoscopic pelvic lymphadenectomy, radical resection of bladder, extracorporeal formation of ileal pouch; extracorporeal implantation of ureters; and laparoscopic urethra-neobladder anastomosis. Erectile nerve sparing procedure was performed for 26 cases. The patients were followed up for 1 - 53 months. RESULTS: The median operation time was 330 min, and the median blood loss was 320 ml. Conversion to open surgery was not necessary in any of the patients. There was no peri-operative mortality. The complication rate was 18.5% (20/108). Surgical margins were tumor free for all cases. The day-time and night-time continence rates were 90.7% and 82.6% respectively in 6 months postoperatively. 10 of the 26 patients undergoing erectile nerve-sparing procedure had potency for intercourse. Follow-up showed 5 cases with local recurrence, 1 case with trocar site seeding and 6 cases with distant metastasis, 8 of the patients died of tumor-related disease and 3 died of diseases not related to tumor. CONCLUSION: Laparoscopic radical cystectomy with extracorporeal formation of orthotopic ileal neobladder is a feasible technique with low morbidity and acceptable neobladder function.

[Compare of laparoscopic and open surgery for radical cystectomy with orthotopic ileal neobladder].

OBJECTIVE: To compare the clinical therapeutic effect and complications of laparoscopic radical cystectomy with orthotopic ileal neobladder (LRC-INB) with open radical cystectomy with orthotopic ileal neobladder (ORC-INB). METHODS: A total of 171 patients were evaluated, including 63 cases with ORC-INB and 108 cases with LRC-INB from June 1994 to May 2007 at our institution. The parameters analyzed included perioperative data, postoperative complications, new bladder function and effect of tumor control. RESULTS: There was no significant difference in demographic characteristics of patients between these 2 groups. The mean operating time was 330 min in the LRC group and 310 min in the ORC group (P > 0.05). The mean blood loss was 320 ml in the LRC group and 1100 ml in ORC group (P < 0.001). The mean oral intake after operation was 2.4 days for LRC group and 4.5 days for ORC group (P < 0.001). No perioperative death was occurred in both groups. The complication rate was 18.5% in LRC group, while 30.0% in ORC group (P < 0.05). Twelve months after operation, the day-time and night-time continence rate were 90.7% and 82.6% for the LRC group, 88.3% and 81.6% for the ORC group respectively (P > 0.05). There was no significant difference of VOL, pressure and residual urine volume (RUV) of neobladder between these 2 groups. Surgical margin were tumor free for 107 cases except one T4 case in laparoscopic group had positive margin (P > 0.05). The mean number of removed lymph node were 12 and 8 in LRC and ORC group respectively (P < 0.05). The 2 years tumor free survival rate of the same stage or grade was no significant different (P > 0.05). CONCLUSIONS: LCR had advantages of less blood loss, shorter oral intake time, less postoperative complications, comparable continent rate and short-term tumor control with ORC. Long-term follow up is needed to confirm the oncological outcome.

A Genomic-clinicopathologic Nomogram for the Preoperative Prediction of Lymph Node Metastasis in Bladder Cancer.

Preoperative lymph node (LN) status is important for the treatment of bladder cancer (BCa). Here, we report a genomic-clinicopathologic nomogram for preoperatively predicting LN metastasis in BCa. In the discovery stage, 325 BCa patients from TCGA were involved and LN-status-related mRNAs were selected. In the training stage, multivariate logistic regression analysis was used to developed a genomic-clinicopathologic nomogram for preoperative LN metastasis prediction in the training set (SYSMH set, n=178). In the validation stage, we validated the nomogram using two independent sample sets (SYSUCC set, n=142; RJH set, n=104) with respect to its discrimination, calibration and clinical usefulness. As results, we identified five LN-status-related mRNAs, including ADRA1D, COL10A1, DKK2, HIST2H3D and MMP11. Then, a genomic classifier was developed to classify patients into high- and low-risk groups in the training set. Furthermore, a nomogram incorporating the five-mRNA-based classifier, image-based LN status, transurethral resection (TUR) T stage, and TUR lymphovascular invasion (LVI) was constructed in the training set, which performed well in the training and validation sets. Decision curve analysis demonstrated the clinical value of our nomogram. Thus, our genomic-clinicopathologic nomogram shows favorable discriminatory ability and may aid in clinical decision-making, especially for cN-patients.

[Screening and characterization of human phage Fab antibody gene to prostate specific membrane antigen].

OBJECTIVE: To screen and characterize the variable region gene about prostate specific membrane antigen (PSMA) of the Chinese Fab fragment, and to establish a new approach to researches on PSMA and prostate gene therapy. METHODS: We used purified PSMA protein as antigen, stuck it on the ELISA plate and scanned the phage Fab fragment antibody library by phage display technology. After five cycles of "absorbing-elution-amplification", we got the Fab fragment phage antibody of PSMA with high antigen binding ability and specificity, and tested it with immunodetection and sequencing. RESULTS: The sequence of Fd fragment was 696 base pairs encoding 232 amino-acid residues, with 98% homological similarity to the human immunoglobulin gamma chain, while the light chain was constructed by 630 base pairs encoding 210 amino-acid residues, with 93% homological similarity to kappa chain. CONCLUSION: Using phage display technology, we obtained the gene sequence of Fab antibody fragment specific to PSMA, and the antibody gene has the classic structural features of immunoglobulin light chain and heavy chain. The coding output of the antibody gene has the specificity and immunological competence to PSMA.

[Identification of response element gene sequence for non-steroid hormone transcription factors for the activation and up-regulation of L-plastin expression in prostate cancer].

OBJECTIVE: To search and identify the non-steroid receptor binding cis-acting elements in the L-plastin promoter in prostate cancer, and the correlative regulation pathway and transcription factors. METHODS: On the basis of construction of the L-plastin promoter luciferase vectors which were removed the steroid hormone receptor AR and ER binding elements, the promoter on the vector was nest-deleted by Exonuclease III and the relative luciferase plasmids were constructed. Transfected these twelve plasmids into prostate cancer cell line LNCaP under dihydrotestosterone-stimulated situation or not and test the intensity of luciferase, then we got the regulation message of every 200 bp part of the promoter in prostate cancer. After the analysis of relative programme, we got the possible regu- lation pathway of non-steroid hormone transcription factors. After removing the possible transcription factors binding site sequence by site-specific mutagenesis, the changes luciferase of activities proved our reasoning. RESULTS: We succeed in segmental deletion of the L-plastin promoter, and constructing the relative plasmids containing part L-plastin promoter on luciferase vector pGL3-basic. After testing the luciferase activities of constructed plasmids, we found the sequence from 206 to 1 of L-plastin promoter had significant luciferase activity. The software TRANSFECT showed that there were binding elements for transcription factors AP-4 at seq-198 to 192 and SP-1 at seq-54 to 41 on the short part promoter (206 to 1). The recombinant plasmids deleted the AP-4 and SP-1 binding elements had lower luciferase activity than the wild-type. CONCLUSION: There are some other non-steroid hormone pathway to regulate the expression of L-plastin except the steroid hormone pathway in prostate cancer. The main binding sites of the non-steroid hormone regulator lies in the sequence from 206 to 1. Transcription factors AP4 and SP-1 may up-regulated the expression of L-plastin by binding these sites.

[Needle laparoscopic varicocelectomy].

OBJECTIVE: We evaluated our method and effects of needle laparoscopic varicocelectomy for the treatment of varicocele. METHODS: 72 patients (105 lateral) diagnosed varicocele were performed laparoscopic varicocelectomy under epidural combined intravenous anesthesia from Feb, 2003 to Apr, 2005. Two 2 mm incisions and one 5 mm incision were made on the midline of lower abdomen, by which two 2 mm trocars and one 5 mm trocar were introduced. Vessel-sealing device (Ligasure) was used to seal the internal spermatic veins. RESULTS: All operations were completed successfully. Mean operation time was 15 minutes unilateral and 21 minutes bilateral. The patients were hospitalized for 3 to 5 days after procedure. Follow-up was scheduled for 6 to 12 months and there was no recurrence. CONCLUSION: Needle laparoscopic varicocelectomy gives favorable effect with minimal invasion, rapid recovery, which is the best choice for the treatment of varicocele.

MicroRNA-155 promotes bladder cancer growth by repressing the tumor suppressor DMTF1.

MicroRNA-155 (miR-155) is dysregulated in human cancers. In this study, we reported that miR-155 was over-expressed in bladder cancer tissues. We found that miR-155 promoted cell proliferation in vitro and tumorigenesis in vivo. MiR-155 directly reduced the expression of the tumor suppressor DMTF1. The expression of DMTF1 was decreased in bladder cancer tissues. Similar to the restoring miR-155 expression, knockdown of DMTF1 promoted cell growth and cell cycle progression, whereas DMTF1 over-expression rescued the effect of miR-155. Moreover, we investigated DMTF1-Arf-p53 pathway and found that DMTF1 worked in both p53-dependent and p53-independent manners. Taken together, our findings suggested that miR-155 functions as a tumor promoter in bladder cancer, which is partially through repressing DMTF1 expression. The identification of miR-155 and its novel target DMTF1 will be valuable in developing diagnostic markers and therapeutic applications for bladder cancer.

Kidney cancer cells secrete IL-8 to activate Akt and promote migration of mesenchymal stem cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent adult stem cells that have the capability of homing to cancer cells. Thus, MSCs play an important role in the development, metastasis, and drug resistance of cancers. The mechanisms underlying the homing of MSCs in kidney cancer are still poorly understood. METHODS: In the present study, enzyme-linked immunosorbent assay was used to measure the level of IL-8 in patients with kidney cancer and in the culture medium of kidney cancer cells. Immunofluorescence staining and reverse transcription polymerase chain reaction were utilized to explore the main receptor for IL-8 in MSCs. Transwell migration assay was performed to measure the migration ability of MSCs and Western blot test was performed to test the activation of signaling pathways. RESULTS: The serum level of IL-8 was markedly increased in patients with kidney cancer, and 2 kidney cancer cell lines were found to secrete IL-8. MSCs had high expression of the IL-8 receptor (CXCR2). Blocking IL-8 or CXCR2 could decrease the migration ability of MSCs. IL-8 could significantly increase Akt phosphorylation in MSCs. CONCLUSIONS: Kidney cancer cells secrete IL-8 to activate the Akt signaling pathway via CXCR2 on MSCs, inducing the migration of MSCs, which may be one of the important mechanisms underlying the homing of MSCs in kidney cancer.

MicroRNA-335 acts as a candidate tumor suppressor in prostate cancer.

MicroRNA-335 (miR-335) acts as a tumor suppressor or a tumor promoter in different human malignancies. However, the involvement of miR-335 in prostate cancer (PCa) is still unclear. The purpose of this study was to investigate the functional and clinical significance of miR-335 in PCa. miR-335 expression in 3 PCa cell lines (LNCaP/DU145/PC3) and in 20 clinical PCa tissues were detected by real-time quantitative reverse transcriptase-PCR compared with corresponding controls. The function of miR-335 was investigated for cell proliferation, invasion and migration in PCa cells transfected with agents containing EGFP-miR-335 expression vector. Additionally, miR-335 expression in 104 clinical PCa tissues was detected by in situ hybridization. Its assocaitions with clinicopathological features and prognosis in patients with PCa were also determined. miR-335 was significantly down-regulated in PCa cell lines than in the normal prostate cell line (P < 0.01). With the similar results in vitro, the reduced expression of miR-335 was also found in human PCa tissues comparing with paired adjacent benign prostate tissues (P < 0.05). Moreover, the increased expression of miR-335 suppressed cell proliferation, invasion and migration of PCa cell lines in vitro. Turning to its clinical significance, the low expression of miR-335 was significantly associated with high Gleason Score (P = 0.04), advanced clinical stage (P = 0.04), and positive metastasis (P = 0.02), but not with prognosis in PCa patients. Our data demonstrated for the first time the inhibitory effect of miR-335 on cell proliferation and invasion for PCa cells. The loss of this microRNA might be associated with clinical progression of PCa patients.

MEK1 and MEK2 differentially regulate human insulin- and insulin glargine-induced human bladder cancer T24 cell proliferation.

BACKGROUND: Increased risk of bladder cancer has been reported in diabetic patients. This study was to investigate the roles of mitogen-activated protein kinase kinase (MEK) 1 and 2 in the regulation of human insulin- and insulin glargine-induced proliferation of human bladder cancer T24 cells. METHODS: In the absence or presence of a selective inhibitor for MEK1 (PD98059) or a specific siRNA for MEK2 (siMEK2), with or without addition of insulin or glargine, T24 cell proliferation was evaluated by cell counting kit (CCK)-8 assay. Protein expression of MEK2, phosphorylation of ERK1/2 and Akt was analyzed by Western blotting. RESULTS: T24 cell proliferation was promoted by PD98059 at 5 - 20 µmol/L, inhibited by siMEK2 at 25 - 100 nmol/L. PD98059 and siMEK2 remarkably reduced phosphorylated ERK1/2. Insulin- and glargine-induced T24 cell proliferation was enhanced by PD98059, suppressed while not blocked by siMEK2. Insulin- and glargine-induced ERK1/2 activation was blocked by PD98059 or siMEK2 treatment, whereas activation of Akt was not affected. CONCLUSION: MEK1 inhibits while MEK2 contributes to normal and human insulin- and insulin glargine-induced human bladder cancer T24 cell proliferation.

Lipopolysaccharide-enhanced early proliferation of insulin secreting NIT-1 cell is associated with nuclear factor-kappaB- mediated inhibition of caspase 3 cleavage.

BACKGROUND: Increased levels of plasma lipopolysaccharide (LPS) have been found in obesity and diabetes patients. This study was to investigate the effect of LPS on pancreatic beta-cell viability and the involvement of caspase 3 in NIT-1 cell line. METHODS: Mouse insulinoma NIT-1 cells were treated with LPS for the indicated time and dose. Cell viability was measured by cell counting kit-8 reagent. Toll-like receptor 4 (TLR4), caspase 3 and cleaved caspase 3 were detected by Western blotting. Insulin was determined by radioimmunoassay (RIA). RESULTS: LPS promoted NIT-1 cell proliferation at 1 µg/ml, peaked at 72 hours of incubation. A reduction in cleavage of caspase 3 was observed upon LPS treatment. Bay11-7082, a specific inhibitor of nuclear factor (NF)-κB, blunted LPS-induced inhibition of caspase 3 cleavage. Reduction in chronic insulin secretion was observed after treatment with LPS at 1 µg/ml for 48 and 72 hours, not for 24 hours. TLR4 protein was upregulated when NIT-1 cells were treated with LPS at 1 µg/ml for 24 hours. CONCLUSIONS: LPS promotes early NIT-1 cell proliferation in association with NF-κB-mediated inhibition of caspase 3 cleavage. LPS exerts a time-dependent inhibitory effect on chronic insulin secretion from NIT-1 cells.

[Detection and clinical significance of circulating tumor cells in peripheral blood of breast cancer patients].

BACKGROUND & OBJECTIVE: Recently, immunocytochemistry and immunomagnetic enrichment have been used in detecting circulating tumor cells (CTCs). This study was designed to investigate the sensitivity and specificity of modified immunomagnetic enrichment of tumor cells in combination with fluorescent immunocytochemistry in detecting CTCs in peripheral blood of patients with breast cancer. METHODS: The sensitivity of this detection method was evaluated by sparking breast carcinoma cell line MCF-7 into normal peripheral blood. Mononuclear cells were isolated from peripheral blood of 52 naive breast cancer patients and 20 healthy female volunteers. Epithelial cell adhesion molecule (EpiCAM) antibody (Ab), covalently bound to magnetic beads, was used to enrich CTCs expressing EpiCAM antigen. CTCs, indicated by positive staining of cytokeratin (CK) 8/18 (green fluorescence), was detected by modified fluorescent immunocytochemistry, and confirmed by DAPI nuclear staining (deep blue). RESULTS: The sensitivity of immunomagnetic enrichment with fluorescent immunocytochemistry was so high that 1 tumor cell in 1x10(7) peripheral blood mononuclear cells could be detected. The specificity of this method was 100%. Positive rate of CTCs was significantly higher in peripheral blood of the patients than in peripheral blood of the healthy volunteers (53.8% vs. 0, P<0.001). The presence of CTCs was correlated positively with clinical stage (P<0.001) and axillary lymph node status (P<0.005), and irrelevant with other prognostic factors (P>0.05). CONCLUSIONS: Modified immunomagnetic enrichment of tumor cells in combination with fluorescent immunocytochemistry is a time-saving, easily-performed, sensitive and specific method for detecting CTCs in peripheral blood of breast cancer patients. The presence of CTCs in peripheral blood correlates positively with clinical stage and axillary lymph node status of breast cancer patients.