Objective To explore the value of lymphatic mapping (LM) and sentinel lymph node(SLN) analysis in laparoscopic colectomy for colon carcinoma. Methods Thirty-two patients with clinically localized colonic neoplasms were subjected to submucosal injection of isosulfan blue dye (0.5-1.0 mL) via a colonoscope during operation. Blue-stained lymphatics were visualized through the laparoscope and followed to the SLN,which was tagged. The colectomy was completed in standard fashion. All lymph nodes were stained by hematoxylin and eosin,and multiple sections of each SLN were examined by immunohistochemical (IHC) staining using cytokeratin antibody. Results At least one SLN was identified laparoscopically in all patients. The SLN accurately predicted the tumor status of the nodal basin in 94% of cases. In 8 cases (25%),an unexpected lymphatic drainage pattern altered the extent of mesenteric resection. The SLN was negative by HE staining in 4 (13%) cases,which were demonstrated positive for micrometastases through immunohistochemical staining. Conclusions SLN mapping during laparoscopic colon resection can alter the margins of resection and in combination with immunohistochemical staining may improve staging,which may more accurately assign patients to prospective protocols.

BACKGROUND:The degradable poly-L-lactic acid (PLLA) biliary-enteric composite stent has been developed. OBJECTIVE:To analyze the solubility and biocompatibility of the degradable PLLA biliary-enteric composite stent. METHODS:Solubility:the PLLA composite stent was implanted into artificial gastric acid to detect the dissolution rate within 12 weeks. Pyrogen test:the PLLA composite stent extracts were injected into the rabbits via ear vein to detect the changes of body temperature. Hemolysis test:the PLLA composite stent extracts, normal saline and distil ed water were added into the rabbit anticoagulant, respectively, to detect the hemolysis ratio. Cytocompatibility test:Caco-2 cel s were respectively cultured in the DMEM medium containing 10%fetal bovine serum, rubber material extracts and the PLLA composite stent extracts, and the cel proliferation was detected at 12, 24, 36 and 48 hours;the lactic dehydrogenase release was detected at 2 days. RESULTS AND CONCLUSION:The PLLA composite stent showed a long stability in vivo, and approximately 80%was dissolved at about 20 weeks. No reactions of pyrogen and henolysis were observed in the pyrogen and hemolysis tests. The PLLA composite stent made no effects on the Caco-2 cel proliferation and lactic dehydrogenase release. In conclusion, the PLLA composite material holds a good solubility and cytocompatibility.

Objective To analyze the mutations of FMS-like tyrosine kinase 3 internal tandem duplication (FLT3/ITD) in bone marrow cells of patients with newly-diagnosed acute myeloid leukemia (AML).Methods The mutations of FLT3/ITD in 96 AML cases were detected by polymerase chain reaction (PCR)and the clinical features of FLT3/ITD positive patients were analyzed.Results FLT3/ITD mutations were identified in 18 patients (18.8 ％),2 cases were M2,11 cases were M3,2 cases were M4,3 cases were Ms.Patients with FLT3/ITD mutations presented higher initial white blood cell count than that of patients without FLT3/ITD mutations [18.0×109/L (3.6×109-137.6×109/L) vs 6.3×109/L (4.5×109-113.0×109/L),t =3.04,P ＜ 0.05].Out of FLT3/ITD positive patients,13/16 (81.3 ％) obtained complete remission and 13 patients remained in first remission in a median follow-up of 10 months(6-15 months).Conclusion The mutations of FLT3/ITD are frequently identified in newly-diagnosed AML patients,patients with FLT3/ITD mutations present high white blood cell count.

Objective To investigate the clinical pathological association of FOXQ1 with colorectal carcinoma (CRC) and colorectal adenoma (CRA). Colorectal mucosa specimens were collected between Jane 2007 and June 2009 in the First hospital of Yunnan prorince, and consisted of CRC (n=76), CRA (n=50) and normal (n=48,≥8 cm apart from cancer) tissues. Methods The expression of FOXQ1 in colorectal mucosa with was detected using immunohistochemistry (SP method). PCR amplification and direct DNA sequencing were used to identify FOXQ1 gene mutations in 23 CRC, 22 CRA and 18 normal specimens. Results There was no expression of FOXQ1 in normal specimens. Aberrant expression of FOXQ1 in either CRC or CRA specimens was significantly higher than in normal colorectal mucosa tissue (76.3% vs 0.0%, 26.0% vs 0.0% respectively, P＜0.01).Aberrant expression rates of FOXQ1 was significantly higher in CRC than that in CRA (76.3% vs 26.0%, P＜0.01). Expression level of FOXQ1 was increase gradually along with the pathological process of colorectal adenoma-carcinoma sequence. In CRC, the aberrant expression of FOXQ1 was not only distributed in nuclear of the tumor cells, but also found in the cytoplasm and the extracellular matrix. For CRC patients, the aberrant expression rates of FOXQ1 in extracellular matrix in Dukes C/D was significantly higher than that in Dukes A/B (61.5% vs 3.3%, 61.5% vs 5.0% respectively,P＜0.01);The aberrant expression of FOXQ1 in extracellular matrix in patient with lymph node metastasis was significantly higher than those without lymph node metastasis (61.5 % vs 4.0 %, P＜0.01). FOXQ1 gene mutation was only identified in one out of 23 CRC specimens, but not found in 22CRA nor 18 normal colorectal mucosa samples. Conclusions Aberrant expression of FOXQ1 may play an important role in the carcinogenesis of CRC and FOXQ1 gene may be involved in the cancer erosion and lymph node metastasis. FOXQ1 mutation is not the primary cause of aberrant FOXQ1 expression in CRC.

Objective The frequency and the anatomic distribution of involved regional nodes in recurrent and locally advanced breast cancer were analyzed, in order to evaluate the rational of conventional regional node radiation technique and provide evidence for target definition of breast cancer . Methods Patients with recurrent or locally advanced breast cancer who were treated in our hospital from August 2003 to December 2009 were included in this study. 111 patients had contrast enhanced chest CT images of the whole regional nodes before treatment. The regional nodes were categorized into 8 anatomical substructures including medial and lateral supraclavicular nodes ( SC-M, SC-L), axilla nodes ( ALN )- Ⅰ , Ⅱ , Ⅲ,infraclavicular nodes (IFN), Rotter's nodes (RN) and internal mammary nodes (IMN). The frequency of involvement and anatomical distribution of the involved nodes on CT images were analyzed. Results A total of 111 patients were enrolled this study and 199 anatomical substructures with involved nodes were identified. The frequency of involvement were :SC-M 33, SC-L 21, ALN- Ⅰ 30, ALN-Ⅱ 25, ALN-Ⅲ + IFN 35, RN 27, IMN 28. Supraclavicular region and axilla were the most frequently involved area (72. 3% ).The average depth of the SC-M and SC-L nodes was 33.48 mm ± 10. 57 mm and 45.62 mm ±20. 45 mm,and 51.5% and 71.4% of the SC-M and SC-L nodes were located more than 3 cm deep from the skin. The axilla nodes were located cranial and caudal to the axillary vein in 5 and 20 locally advanced breast cancer patients and in 64 and 28 patients who received prior axillary dissection. The majority of involved IMN was located within the first 3 intercostal spaces (26/28). The average distance between the center of involved IMN and chest skin was 24. 23 mm ± 10. 28 mm. The average distance between the center of involved IMN and midline of the body was 29. 38 mm ±6. 7 mm. The center of involved IMN was 6.19 mm ±5.73 mm lateral and 5.73 mm ± 4. 56 mm posterior to the internal mammary vessels. Conclusions Conventional field design is unlikely to provide sufficient dose to the entire risk region because of individual differences.Individualized treatment planning based on CT would become feasible with increasing knowledge of natural risk of nodal involvement.

BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.

Objective To explore the clinical value of early diagnosis of atherosclerosis in patients with systemic lupus erythematosus (SLE) using vascular echo-tracking technique and to detect changes of elastieity of carotid artery quantitatively in SLE patients.Methods Fifry patients with SLE were divided into SLE1 group(duration≤1 year),and SLE2 group(duration>1 year)based on different course.An ultrasonic echo-tracking method was used to measure patients'pressure strain elastic modulus (Ep),the stiffness constant(β),arterial compliance(AC),augmentation index(AI),pulse wave velocity (PWVβ) and intimamedia thickness(IMT)of the common carotid arteries in 50 patients with SLE and in 25 healthy control subjects.Results Among carotid artery elasticity indicators of three groups,there was no significant difference in AI(P>0.05).Ep,β,PWVβ parameters of SLE1 group and SLE2 group were statistically higher than that of the control group[Ep of SLE1 group,SLE2 group,control group was (69±20),(103±40),(48±18)kPa respectively;β was 5.2±1.9,8.0±3.1,4.2±1.3 respectively;PWVβ was 5.2±0.7,6.3±1.1,4.5±0.7]respectively,but AC(AC of SLE1 group,SLE2 group,control group was(1.1±0.3),(0.8±0.3),(1.2±0.6)mm2/k respectively]lower than the controls(P<0.01).Ep,β,PWVβ in SLE2 group was significantly increased compared with SLE1 group,but AC was decreased (P<0.01).Conclusion The application of echo-tracking technology can be used to diagnose early atherosclerosis.Complications of cardiovascular disease in SLE have high clinical value.

Objective To assess set-up errors measured with kilovoltage cone-beam CT (KV-CBCT), and the impact of online corrections on margins required to account for set-up variability during IMRT for patients with prostate cancer. Methods Seven patients with prostate cancer undergoing IMRT were enrolled onto the study. The KV-CBCT scans were acquired at least twice weekly. After initial set-up using the skin marks, a CBCT scan was acquired and registered with the planning CT to determine the setup errors using an auto grey-scale registration software. Corrections would be made by moving the table if the setup errors were considered clinically significant ( i. e. , ＞ 2 mm). A second CBCT scan was acquired immediately after the corrections to evaluate the residual error. PTV margins were derived to account for the measured set-up errors and residual errors determined for this group of patients. Results 197 KV-CBCT images in total were acquired. The random and systematic positioning errors and calculated PTV margins without correction in mm were:a) Lateral 3. 1,2. 1,9. 3;b) Longitudinal 1.5, 1.8, 5. 1 ;c) Vertical 4. 2,3.7, 13.0. The random and systematic positioning errors and calculated PTV margin with correction in mm were:a) Lateral 1.1,0. 9, 3.4;b) Longitudinal 0. 7, 1.1, 2. 5;c) Vertical 1.1, 1.3, 3.7. Conclusions With the guidance of online KV-CBCT, set-up errors could be reduced significantly for patients with prostate cancer receiving IMRT. The margin required after online CBCT correction for the patients enrolled in the study would be appoximatively 3-4 mm.

Objective To investigate the expression pattern of transcription factor Olig 2 in cuprizone-induced mouse model of acute demyelination .Methods C57BL/6 mice were fed with 0.2%cuprizone to induce acute demyelina-tion.Immunofluorescence and qRT-PCR were used, and Olig2, MBP and GFAP were detected in the brain tissues of con-trol group and cuprizone-treated groups for 6 weeks and recovery for 2 weeks.Results Severe demyelination occurred in the corpus callosum following 6-weeks exposure to cuprizone , while remyelination was detected in the white matter after the mice were given diet without cuprizone .In the normal mice , Olig2 was expressed in a low level , while the experessions of Olig2 and GFAP were significantly increased , and Olig2 +/GFAP+cells were detected after demyelination .But the expres-sion of MBP was below the normal level with demyelination .After recovery for 2 weeks, the experession of Olig2 was lower, but the experessions of MBP and GFAP were increased .Conclusions Olig2 may play an important role in the glial differ-entiation from neural progenitor cells into active astrocytes , and in the glial scar formation .

OBJECTIVE:To research the mechanism of in vitro permeation of phillyrin through blood-brain barrier. METH-ODS:After Madin-Darby canine kidney epithelial cells transfected with colorectal cancer MDR1 gene (MDCK-MDR1) were cul-tured with phillyrin solution of 0(negative control),10,25,50,75 and 100μg/ml for 24 h,cell viability was determined by resa-zurin method and cell survival rate was calculated. After MDCK-MDR1 cells were cultured with phillyrin solution of 10,25,50, 75 and 100 μg/ml for 10 min,the content of phillyrin in the cells was determined,and concentration-uptake rate curve was drawn. Following 3 h culture of MDCK-MDR1 cells with phillyrin solution of 0 (negative control),50 and 100 μg/ml,the structure of cell tight junction protein was observed under the inverted microscope. RESULTS:Compared to the negative control group,after 24 h cell culture with phillyrin solution of 10-100 μg/ml,no obvious change in cell survival rate occurred. MDCK-MDR1 cells cultured with the phillyrin at a mass concentration of 10-100 μg/ml demonstrated a nonlinear relationship with concentration of phillyrin and a gradual saturation trend. After the cells were cultured with phillyrin of 50 and 100 μg/ml for 3 h,cell tight junction protein was intact. CONCLUSIONS:The absorption of phillyrin through the simulated blood-brain barrier may be in the form of passive transportation combined with active transportation,the concertration has effect on cell tight junction protein.