Background and purpose:Majority of patients with B cell lymphoma often achieve complete clinical remission after systemic treatment, but half of the patients ultimately relapse. The residual neoplastic cells, commonly called ‘minimal residual disease’ (MRD), are thought to be the source of relapse. But not all of the patients whose results of IgH rearrangement were positive had relapse or distant involvement. It was thought that the patients whose IgH rearrangement was positive relapsed or not may be associated with the quantity of IgH rearrangement. The study tried to investigate the feasibility and clinical significance of detecting immunoglobulin heavy chain gene in DLBCL by SYBR Green RT-FQ-PCR. Methods:Fifty-seven bone marrow specimens from 44 patients diagnosed with DLBCL were used to detect IgH-R. Namalwa cell line and U-937 cell line were used for positive and negative control respectively. The ?-actin gene was chosen as inter control. DNA was isolated by phenol-chloroform-isoamyl alcohol and then was amplified by SYBR Green RT-FQ-PCR targeting the IgH-R CDR Ⅲ.Results:Melting curve analysis could confirm the specificity of IgH-R. The positive rate detected by RT-FQ-PCR was 63.2%. The positive results of IgH/?-actin were between 0.01 and 4131.69, and the median was 0.42. There was a significant difference between stage Ⅰ/Ⅱ and stage Ⅲ/Ⅳ in IgH-R quantity (P=0.018). Nonparametric test showed that there was a significant difference between patients with normal LDH and patients with elevated LDH (P=0.046).Conclusions:SYBR Green RT-FQ-PCR is a valuable, feasible and sensitive tool to detect IgH rearrangement in DLBCL. Detecting IgH-R using RT-FQ-PCR can help staging more accurately.

Objective To investigate the methylation status of the CpG island of tumor suppressor gene PCDH 8 and its clinical significance in bladder cancer tissues .Methods 79 cases of primary bladder transitional cell carcinoma and 20 cases of normal blad-der mucosa tissue were collected ,and then the promoter methylation status of PCDH8 gene was examined by methylation specific PCR (MSP) ,and correlated with clinical pathological data for statistical analysis .Results We found that no PCDH8 gene methyla-tion was detected in 20 normal bladder mucous tissues ,while PCDH8 promoter methylation was found in 44 cases of total 79 prima-ry bladder transitional cell carcinoma tissues ,the methylation rate was 55 .7% ,and the difference was statistical significant between normal bladder mucous group and bladder cancer group (P<0 .01) .The promoter methylation of PCDH8 gene in bladder transi-tional cell carcinoma tissues did not correlate with patient′s age ,gender ,tumor number (P>0 .05) ,the methylation rate of PCDH8 gene in tumors whose diameter more than 3 cm was 72 .7% ,while the methylation rate of PCDH8 gene in tumors whose diameter less than 3 cm was 43 .5% ,and the difference was significant(P< 0 .05) .The methylation rate of PCDH8 gene in the papillary tumor was 48 .2% ,while the methylation rate of PCDH8 gene in the unpapillary tumor was 73 .9% ,and the difference was signifi-cant(P<0 .05) .The methylation rate of PCDH8 gene in recurrent tumors was 71 .1% ,while the methylation rate of PCDH8 gene in primary tumors was 35 .3% ,and the difference was significant(P<0 .05) .The methylation rate of PCDH8 gene in tumors with G1 ,G2 phase was 43 .4% ,while the methylation rate of PCDH8 gene in tumors with G3 was 80 .8% ,and the difference was signifi-cant(P<0 .05) .The methylation rate of PCDH8 gene in the tumors with Ta T1 phase was 43 .7% ,while the methylation rate of PCDH8 gene in tumors with T2 T4 was 74 .2% ,and the difference was significant(P<0 .05) .Our result suggested that PCDH8 gene methylation was associated with tumor growth ,morphology ,recurrence ,poor differentiation and tumor invasion (P<0 .05) . Conclusion The promoter methylation of tumor suppressor gene PCDH8 is closely correlated with the occurrence and development of primary bladder transitional cell carcinoma .The promoter methylation of PCDH8 gene could be used as molecular markers of ear-ly diagnosis ,monitoring and prognosis biomarker in bladder cancer .

AIM: To investigate the effect of 5 - fluorouracil ( 5 - FU ) on the expression of the stem cell marker CD133 on colon cancer stem cells. METHODS:CD133 expression on several colon cancer cell lines was detected by flow cytometry. The CD133 positive cells from DLD1 cells were separated by the method of magnetic activated cell separation. Colony assay was used to measure self - renew ability and MTS assay was used to detect the sensitivity to 5 - FU after separation. After 5 - FU treatment, the change of CD133 mRNA level was measured by qPCR. RESULTS: CD133 expression on the surface of colon cacner cell lines DLD1, HT29, SW480, HCT116, Lovo, RKO was 30.20% , 82.00% , 0.34% , 91.80% , 85.30% , 0.28% respectively. DLD1 cells had two obvious populations according to CD133 expression. CD133 positive cells were separated from DLD1 cells, the positive purity was 87.21% ±5.33% and the negative purity was 84.30% ±4.65%. CD133 positive cells formed more colonies with limited dilution colony assay (46.33% ±4.44% vs 31.00% ±2.00% , P <0.05). CD133 positive cells were less sensitive to 5 - FU compared to CD133 negative cells(20% less, P <0.01). 5 - FU at concentration of 1 mg/L upregulated CD133 mRNA expression in both DLD1 and HT29 cells, the relative quantity was increased from 1 to 1.684 ±0.012(P <0.01 )and 30.702 ±0.280 to 49.379 ±0.460(P <0.01) in HT29 and DLD1, respectively. CONCLUSION: Compared to CD133 negative cells, CD133 positive cells show more ability to form colonies in vitro, and are less sensitive to 5 - FU. 5 - FU upregulats the mRNA expression of CD133, resulting in the CD133 colon cancer stem cells enrichment during 5 - FU treatment.

Objective To reassess the treatment of Bosniak category Ⅱ-Ⅲ renal cyst.Methods Sixty-eight cases of Bosniak category Ⅱ-Ⅲ renal cyst were operated from 2005 to 2008 in our institute.The average patient age was 40 years with the average renal cyst diameter of 5.1 cm.There were 37 left lesions and 31 right lesions with 31 cases of category Ⅱ (including 13 cases of category Ⅱ F) and 37 cases of category Ⅲ.Renal unroofing were performed in 49 cases with 9 cases found malignant and followed by radical nephrectomy,19 cases were diagnosed as malignancy preoperatively and nephron sparing surgery or radical nephrectomy were performed in these cases.Results Malignant lesions were found in 21 cases and benign lesions were found in 47 cases.The malignant rate was 9.7％ in category Ⅱ (15.4％ in category Ⅱ F),48.6％ in category Ⅲ.Nine cases were diagnosed benign preoperatively but confirmed malignancy after operation; 7 cases were diagnosed malignancy preoperatively but confirmed benign after operation.Forty-two cases were followed up for 8-65 months with 15 cases of malignancy and 27 cases of benign lesions.The recurrence of renal cyst occurred in 6 cases of benign lesions and recurrence and metastasis occurred in 3 cases of malignancy in 1-5 years.Conclusions It is difficult to draw a therapeutic principle for Bosniak category Ⅱ-Ⅲ renal cyst because of the uncertainty of the lesion.Partial nephrectomy or renal cystectomy is a good choice in the treatment of Bosniak category Ⅱ-Ⅲ renal cysts.

Objective To investigate the role of radiotherapy (RT) and prognostic factors in the combined modality treatment (CMT) of patients with stage ⅠE-ⅡE extranodal nasal type NK/T-cell lym-phoma. Methods From Dec. 1990 to Dec. 2006,177 patients who were diagnosed and treated in our hos-pital were retrospectively analyzed,induding 37 received chemotherapy (CT) alone ( median 4 cycles), 128 received CT (median 3 cycles) followed by RT (median 52 Gy) ,6 received RT alone (median 58 Gy) and 6 received RT ( median 54 Gy) followed by CT ( median 5 cycles). Results The overall response ( CR + PR) rate after initial CT was 60.8% compared with 83.8% after RT ( x2 = 28.63, P < 0.01 ). The 5-year overall survival (OS) and progress-free survival (PFS) rates were 46.2% and 36.8% ,respectively. The lo-cal control rates were 80.9% for RT ( alone or with CMT) and 50.0% for CT alone (x2 = 14.39, P < 0.01 ), and corresponding 5-year OS and PFS were 53.4% vs. 18.3 % ( x2 = 23.38, P < 0.01 ) and 45.0% vs. 10.9% (x2 =23.46,P <0.01 ),respectively. Compared with CT alone,the following definitive RT for patients who achieved response or not after initial CT significantly improved the local control [83.5%, 76.2% vs. 50.0% (x2 = 14.13,P <0.01;x2 =5.78,P <0.01)] and 5-year OS[56.2%,48.6% vs. 18.3%(x2 =28.87,P <0. 05;x2 =4.80,P <0.05)]. Concinsions Compared with CT alone, RT a-chieves better tumor response, local control and survival of patients not only with tumor response but also with local progression after CT. Definitive RT should be the reasonable choice of treatment for early stage extran-odal nasal type NK/T-cell lymphoma.

ObjectiveTo study the risk factors of renal artery pseudoaneurysm (RAP) following partial nephrectomy.MethodsOpen partial nephrectomy was performed on a total of 464 cases of renal cell cancer from July 2003 to May 2010. Five patients ( 1.1％ ) had postoperative hemorrhage from RAP.The surgery technique of the open partial nephrectomy, the clinical presentation, imaging findings and treatment of RAP were reviewed. The anatomical characteristics of these five renal tumors on enhanced CT were quantified using the R.E.N.A.L. Nephrometry Score System.ResultsAll five cases were male, two had tumors on the left side and three on the right side. Median tumor size was 3.6 cm ( range from 2.5 to 5 cm; Radius score 1 - 2). Four tumors were exophytic of these, three had a major endophytic component (≥50％) deep in the parenchyma (Exophytic/endophytic score 2 ), one was entirely endophytic (score 3 ). The distance of all the tumors to the collecting system was ≤4 mm ( Nearness score 3 ). Four of the five tumors were across the polar line and/or renal axial midline ( Location score 3 ). The other tumor was located under the lower pole ( Location score 1 ) but close to the renal hilar. All patients presented with delayed gross haematuria and decreasing hemoglobin occurred on mean postoperative day 12 (3 -23 day). Four patients complained of flank pain, two of which had signs of hypovolemia requiring blood transfusion. The diagnosis was confirmed by the contrast medium-enhanced CT and selective angiography, and RAP was found most commonly arising from the segmental branch of renal artery. Superselective microcoil angioembolization was successfully performed in four cases, once in three cases and twice in the remaining case. The procedure failed in one patient and a nephrectomy was done. At a mean follow-up of 21 months (12 -30) , all patients had normal renal function without evidence of recurrence.ConclusionsRAP should be considered in all patients who had delayed hematuria after partial nephrectomy. A central, deep tumor and its relationship to the segmental branch of renal artery could be an important risk factor for this complication. Choosing the case properly for partial nephrectomy and suturing the transected vessels and the defect of parenchymal correctly could reduce occurrence of this serious complication. Early use of selective angioembolization could be a primary choice of treatment.

Objective To evaluate the application significance of Bosniak renal cyst classification. Methods The operations of 497 cases of cystic renal masses were performed from 2002 to 2007 in our hospital.The average age of these cases was 42-years old,the average diameter of renal cysts was 5.4 cm. There were 212 left lesions,265 right and 20 bilateral.According to Bosniak classification,there were 339 cases of category I,49 cases of category Ⅱ(including 23 cases of category ⅡF),44 cases of category Ⅲ and 65 cases of category Ⅳ.Renal cystectomy was performed in 372 cases when nephron sparing surgery performed in 51 eases and radical nephrectomy performed in 74 cases.Results Malignant tumors were found in 89 cases and benign lesions were found in 408 cases.The malignant rate was 0.9% in category I,10.2% in category Ⅱ(13.O%in category ⅡF),52.3% in category Ⅲ,89.2% in category Ⅳ.There was significant difference among the 4 groups,but there was no significant difference between category ⅡF and non-Ⅱ F of category Ⅱ.403 cases were followed up for 1-5 years with 75 cases of malignancy and 328 cases of benign lesions.Recurrence or metastasis was found in 9 malignant cases.Conclusions Careful follow-up should be performed in category I and Ⅱ,especially in the cases of Ⅱ F category and the eases with changes of diameter or morphology.It is definite that surgery should be performed in the cases of renal cyst with category Ⅲ or Ⅳ for their high malignant rate.

Objective To discuss the etiology and management of hemorrhage after partial nephrectomy because of renal cancer. Methods Of the 382 cases performed partial nephrectomy during a 10-year period, 5 patients(1.3%) had postoperative hemorrhage because of a renal artery pseudoaneurysm. The ages of these patients were from 42 to 63 years. All the tumors were malignant with an average diameter of 2. 8 cm. The renal pedicle was blocked in 2 cases with tumor size larger than 3 cm. The kidney was reconstructed with interrupted 0 chromic sutures in "8"or "U" style. Delayed postoperative hemorrhage occurred from 6 days to 3 months. The blood loss was from 1000 to 4500mi. Shock or lower blood pressure occurred in 3 patients. Nephrectomy was performed in 1 patient and selective angiographic embolization was performed in other 4 cases. Results Angiography revealed renal artery pseudoaneurysm in all 5 cases at the third (4 cases) or fourth branch(1 patient).Percutaneous embolization was successful in 4 patients. Four cases were followed up for 2 to 9 years without complication. Conclusions Delayed postoperative hemorrhage following partial nephrectomy is an uncommon but potentially life-threatening complication. Super-selective angiographic embolization could be the first choice of treatment.

Objective To discuss the treatment of synchronous bilateral renal cell carcinoma.Methods Fourteen cases of synchronous bilateral renal cell carcinoma with 12 males and 2 females were treated from 1994 to 2005.Lumbago occurred in 4 cases and hematuria in 2 cases.All the cases were diagnosed with the CT scan,IVU,US and/or MR.Six cases received bilateral operation simultaneously,8 cases received sequential operations.Radical nephrectomy on one side and partial nephrectomy on the opposite side was conducted in 8 patients.Partial nephrectomy on both sides was performed for 6 patients. Results Twenty-two operations were performed in 14 cases.The average operative time was(263±52)min in simultaneous operation and(155±46)min in others.The length of hospital stay was(11.5±2.1)d in simultaneous operation and(7.4±1.2)d in staging operatiom Ten cases were followed up for 6-41 months.The renal function remained good in 7 patients.The SCr increased in 3 cases with no need for dialysis.One case died 25 months later with metastasis of lung.One case with local recurrence died 38 months later.One case died of cerebral hemorrhage.Conclusions The renal function and treating tumor must be considered in managing synchronous bilateral renal cell carcinoma.The geheral condition of patient and the position of tumor should be evaluated in order to make a treatment plan.

Objective To observe the protection and distribution of bone marrow mesenchymal stem cells (MSCs) by distinct intravenous infusion time on renal ischemia reperfusion injury (IRI) in rats.Methods We used unilateral nephrectomy and contralateral vascular occlusion method to establish renal IRI model in rats.The experimental groups which received 2 × 106 MSCs infusion through the tail vein,were subsequently divided into 3 subgroups:2 h pre-reperfusion (PreOp,n =16),immediately after reperfusion (Op,n =16),6 h post-reperfusion (PostOp,n - 16).The control groups included sham operation group (n =16) and ischemia group (n =16).Chemotaxis of DAPI-labeled MSCs was detected 6 h after administration in the IR kidney.Renal function was detected at 6,24,and 48 h respectively after operation. Forty eight h after operation,the renal tissues were harvested to observe the pathological changes by HE staining and the tubular epithelial cell apoptosis via TUNEL assay.Results MSCs were found in the experimental groups after IR in the kidney,most in PostOp group.Twenty-four and 48 h after reperfusion,there was no significant difference in Cr and BUN between the experimental groups and sham operation group (P＞0.05),but the levels of Cr and BUN in the experimental groups were significantly lower than in the IR group (P＜ 0.05). As compared with IR group,the renal pathological injury was alleviated,the number of apoptotic cells was decreased in the experimental group,most significantly in PostOp group (P＜0.05).Conclusion MSCs can reduce the inflammatory response and inhibit renal tubular cell apoptosis in rat renal IRI.Post-reperfusion administration of MSCs leads to the best chemotaxis efficiency and protection.