Objective To investigate the clinicopathological features of renal cell carcinoma (RCC).Methods From December 1956 to August 2012,the clinicopathological features of RCC were studied in 705 cases and related literatures were reviewed.Results The diameter of RCC ranged from 0.6 to 18.0 cm,which the average size was 4.6 cm.The proportions of the clear cell,papillary,multilocular clear cell,chromophobe and unclassified histologic subtype were 88.9％ (627/705),4.1％ (29/705),3.3％ (23/705),1.3％ (9/705) and 2.4％ (17/705),respectively.According to the Fuhrman grading system,the proportions of grade 1,2,3,4 were 19.0％ (116/612),58.3％ (357/612),18.1％ (111/612)and 4.6％ (28/612),respectively.The rates of invasion into the renal pelvis,perirenal fat and vascular were 10.9％ (66/603),10.6％ (64/603) and 4.8％ (29/603),respectively.Of 705 cases,464 (76.6％)cases were in T1,65 (10.7％) cases in T2,73 (12.0％) cases in T3,and 4 (0.7％) cases in T4.As to the lymph node and distant metastasis,the rate was 2.8％ (17/606) and 3.5％ (21/606).The percentages of stage Ⅰ,Ⅱ,Ⅲ and Ⅳ RCC were 74.3％ (450/606),9.9％ (60/606),11.7％ (71/606) and 4.1％(25/606),respectively.The 3-,5-,10-and 15-year disease-specific survival rate for RCC was 92.8％,86.9％,76.8％ and 55.5％,respectively.To those patients with clear cell RCC,the disease-specific survival at the same time point was 92.8％,88.1％,77.4％ and 55.4％,respectively.Multivariate analysis showed that the stage was the only independent prognostic factor for RCC.Conclusions Tumor stage of RCC is the independent prognostic factor for disease-specific survival.The evaluation of renal sinus invasion and lymph node should be noted in the diagnosis of RCC.

Objective To evaluate the role of P2X3 receptors in dorsal root ganglion in development of incisional pain in rats.Methods Twenty-four healthy male SD rats weighing 200-220 g were randomly divided into 3 groups(n = 8 each): control group(group C),incisional pain(group IP)and P2X3 receptor antagonist + IP group(group A).In group IP and A,a 1 cm longitudinal incision was made in the plantar surface of left hindpaw according to the method described by Brennan et al.in isoflurane-anesthetized rats.P2X3 receptor antagonist TNP-ATP 200 nmol was injected into the plantar surface of left hindpaw 30 min after plantar incision was made in group A,while equal volume of normal saline was given instead of TNP-ATP in group C and IP.The behavior of the hindpaw of the rats were assessed using cumulative pain score within 1 h after injection.The animals were sacri ficed 2 h after injection and the dorsal root ganglion was removed for determination of P2X3 receptor expression and intracellular Ca2+ concentrations.ResultsThe cumulative pain scores,P2X3 receptor expression and Ca2 + concentrations were significantly higher in group IP and A than in group C(P ＜ 0.05).The cumulative pain scores,P2X3 receptor expression and Ca2+ concentrations were significantly lower in group A than in group IP(P ＜0.05).Conclusion P2X3 receptors in dorsal root ganglion is involved in the development of incisional pain through increasing intracellular Ca2+ concentrations in rats.

Objective:To study the effects of rat interleukin-10 (rIL-10) gene treatment on the expression of collagen , matrix metalloproteinase 13(MMP13) and their specific inhibitors the tissue inhibitor of metalloproteinase 1(TIMP1) in porcine serum in-duced liver fibrosis rats then to explore the anti-fibrotic effect of rL-10.Methods:Thirty SD rats were divided into normal control and fibrosis model group.Normal control group (group C) was intraperitoneally injected with 0.5 ml normal sodium twice a week for 8 week, while the fibrosis model group was injected with equal volume of pig serum for 8 week.At the beginning of the 5th week, fibrosis model group was further randomly divided into a fibrosis model subgroup ( group M ) , rIL-10 gene treatment subgroup ( group I ) and empty vector control subgroup(group P).Rats in group C and M were injected with Ringer’s solution as a reagent control via the tail vein weekly, rats in group I were injected with the rIL-10 plasmid pcDNA3-rIL-10, and rats in group P were injected with empty vector pcDNA3.All rats were sacrificed at the end of 8th week, and the liver tissue samples were collected to observe deposition of collegan in liver tissue by sirius red staining and detected the expression of MMP 13 and TIMP1 in the liver tissue by SP immunohistochemistry .Re-sults:Sirius red staining showed that the area of the collegan deposition was dramatically increased in fibrosis model subgroup and emp -ty vector control subgroup compared with the normal control group , and the area of the collagen deposition was dramatically decreased in rIL-10 gene treatment subgroup compared with the fibrosis model and empty vector control subgroup .Immunohistochemistry analysis showed that the expression of MMP 13 and TIMP1 in fibrosis model subgroup and empty vector control subgroup was significantly higher than the normal control group , but compared with normal control group , expression of MMP13 was significantly increased and expres-sion of TIMP1 was significantly decreased in rIL-10 gene treatment subgroup .Compared with fibrosis model subgroup and empty vector control subgroup, the expression of MMP13 and TIMP1 was dramatically decreased in rIL-10 gene treatment subgroup.Conclusion:rIL-10 gene treatment attenuates the area of collagen deposition in liver fibrosis rats associated with downregulation of TIMP 1.

Objective To investigate the effects of Inula Britannica on myocardial caspase-3 and cytochrome c ( cyt c) following overtraining-induced acute myocardial injury in rats.Methods Forty-eight male Wistar rats weighing 200-220 g were randomly divided into 3 groups:group control (group C,n =8) ; group exhausting swim (group E,n =24) and group Inula Britannica (group IB,n =16).The animal model of overtraining-indnced acute myocardial injury was developed by exhausting swim.The animals were forced to swim until they were exhausted.The animals sank to the bottom and no righting reflex or escape response was elicited when they were taken out of water in groups E and IB.In group IB oral Inula Britannica 25 ml/kg was given 24 h and immediately before overtraining.Blood samples were taken from inferior vena cava immediately and at 6,24 h after overtraining in group E and at 6,24 h after overtraining in group IB for determination of serum cardiac troponin I (cTnI) concentration (by ELISA).The animals were sacrificed after blood sampling and myocardial specimens were obtained for microscopic examination and determination of caspase-3 and cyt c expression (by immuno-histochemistry).Results Overtraining significantly increased serum cTnI concentration and up-regulated myocardial caspase-3 and cyt c expression in group E as compared with group C.Oral Inula Britannica significantly attenuated overtraining-induced increase in serum cTnI concentration and myocardial caspase-3 and cyt c expression in group IB as compared with group E.Conclusion Inula Britannica can reduce overtraining-induced acute myocardial injury by down-regulating caspase-3 and cyt c expression.

Aim To establish a method of hydrodynamic-based transfection(HBT) and provide a means in rats of the gene therapy of liver diseases,which allowed an efficient expression of rIL-10 gene in rat liver.Methods Using rIL-10 gene as a reporter gene,different volumes and doses of plasmid DNA solutions were rapidly injected into rat tail vein,then the serum and the tissue of liver,kidney,lung,spleen and heart in different time were collected and the expression of rIL-10 was detected by the methods of RT-PCR,ELISA and immunochemistry.Results Using rIL-10 gene as a reporter gene,the results demonstrated that an efficient transfer and expression of rIL-10 in rat liver could be achieved by a rapid injection of a large volume of rIL-10 DNA solution into rat via tail vein.Maintaining a stable expression of rIL-10 in serum could be assessed by repeated administration.Conclusion The HBT was a simple,convenient and efficient method of gene transfer and expression in rats,which could be used as an effective means to study further gene therapy of rIL-10 in liver diseases.

Objective To investigate the clinical value of clinical application in diagnosis of prostate cancer (PCa) by prostate biopsy guided by rectal ultrasound contrast guided biopsy.Methods One hundred and ninety-eight patients with prostate in Punan Hospital of Pudong New District of Shanghai were investigated.According to different detection methods, the research objects were divided into two groups, the patients were performed with the ultrasound contrast guided biopsy as the imaging group (n =96), the patients with Doppler ultrasound guided biopsy as the ultrasonic group(n =102).The puncture Results were compared with pathological diagnosis.The positive rate of PCa and number of puncture needle were compared with the two puncture methods.The value of the application of the prostate biopsy guided by rectal ultrasound in diagnosis of prostate cancer was evaluated.Result One hundred and ninety-eight patients were all received pathological diagnosis,78 cases benign lesions, 120 cases were diagnosed as PCa.Thirty-six cases of benign lesions were confirmed by pathological biopsy, 60 case PCa.There were 42 cases of benign lesions in ultrasonic group, 60 case PCa.The positive rate of PCa in the imaging groupwas 62.5％ (60/96), the ultrasonic group was 58.82％ (60/102).There was no difference in Pca positive rate between the ultrasound group and the contrast group(x2 =0.104, P=0.747).The positive number of Pca in the imaging group was 28.50％ (17/60), the difference was statistically significant higher than that of the common group(18.80％ (11/60), P =0.001).The average of the patients in the imaging group was 8.19 needle,less than the ultrasonic group per capita 11.31 needle less.When f/tPSA less than or equal to 0.15, Pca positive rate of the contrast group was 46.55％ (27/55), higher than the ultrasonic group(9.30％ (4/43)), the difference was statistically significant (P =0.001);when f/tPSA more than 0.15, the positive rate of Pca in the contrast group was 92.11％ (35/38), less than the ultrasound group (94.92 (56/59)), the difference was not statistically significant (P =0.89).Anal pain, hematuria and hematochezia in contrast group (7.29％ (7/96), 2.08％ (2/96), 10.42％ (10/96)) were significantly less than the ultrasound group(22.55％ (23/102), 8.82％ (9/102), 23.53％ (24/102)), the difference was statistically significant (P =0.003,0.039,0.014).Conclusion Under the guidance of ultrasound contrast, rectal biopsy has important diagnostic value for prostate cancer.Under the premise of reducing the number of puncture needle,can improvethe positive rate of Pca, reduce the pain of patients and the occurrence of complications after puncture.When f/tPSAless than or equal to 0.15,puncture positive rate in contrast group is higher than the ultrasonic group, puncture effect is better.

Objective 17β-estradiol is known to have a neuroprotective effect.The aim of this study was to investigate the effects of 17β-estradiol on propofol-induced neuroapoptosis in primarily cultured cortical neurons. Methods Rat cortical neurons were primarily cultured for 7 days and randomly divided into groups A ( vehicle control) , B, and C, treated with equal volume of 20%intralipid, 500 μmol/L propofol, and 500 μmol/L propofol +0.1 μmol/L 17β-estradiol, respectively.At 12 hours after treatment, the morphology of the neurons was observed under the microscope, their survival rate calculated by MTT, their apoptosis was deter-mined by FCM assay, and their mitochondrial membrane potential measured by fluorescent dye rhodamine 123. Results Compared with group A, group B showed a significantly reduced number of neurons, lack of 3-dimensional appearance, unclear contour, and fractured neuron axons, but a remarkable improvement was observed in the propofol-induced morphological damage in group C.The survival rate of the neurons and the mitochondrial membrane potential were markedly decreased in group B ([52.3 ±5.2]% and [59.1 ± 5.3]%) as compared with groups A ( [99.9 ±3.6]%and [99.6 ± 5.8]%) and C ([90.1 ±7.2]%and [89.2 ±7.1]%) (both P<0.01 ) , while the rate of neuroapoptosis significantly increased in group B ([43.4 ±4.6]%) in comparison with A ([3.1 ±0.2]%) and C ([22.3 ±3.2]%) (both P<0.01). Conclusion 17β-es-tradiol can protect against propofol-induced apoptosis of primarily cul-tured neurons by inhibiting the reduction of their mitochondrial membrane potential.

Objective To explore the relationship between abnormal copy number of mitochondrial DNA (mtDNA) and prognosis of the patients in colorectal cancer (CRC).Methods A total of 60 cases of CRC and corresponding adjacent noncancerous tissue specimens were selected.Genomic DNA was extracted.The mitochondrial ND1 gene was detected by quantitative real-time polymerase chain reaction and β-actin was set as internal control.And then the copy number of mtDNA was caculated and analyzed by t-test.The prognosis of patients was determined by Kaplan-Meier survival analysis.Results The mean mtDNA copy number of CRC tissue was 108.60±20.11,while that of the corresponding adjacent noncancerous tissues was 153.68±25.72,the former was significantly lower than that of the latter (t=10.69,P＜0.01).The rate of low mtDNA copy number in case with positive lymph nodes metastasis was higher than that in cases with negative lymph nodes metastasis (x2 =4.022,P＜0.05),however there were no obvious correlation with gender,age,pathological type,TNM stage.The survival rate of high mtDNA copy number group was higher than that of low mtDNA copy number group,however there was no statistical significance (P＞ 0.05).Conclusions mtDNA copy number of CRC was significantly lower than that of the adjacent noncancerous tissues.However the change of copy number was not correlated to the prognosis.

AIM: To construct the replicative defecient adenovirus Ad-Runx3 expressing Runx3, and to express it in U251 malignant glioblastoma cells. METHODS: The runx3 gene with a flag tag was amplified by PCR using pCMV5-AML2 as a template, and was confirmed by DNA sequencing. The adenovirus shuttle vector pShuttle-CMVRunx3 was constructed by introducing runx3 DNA fragment into the sites of Kpn Ⅰ and Xho Ⅰ of pShuttle-CMV vector. This recombinant plasmid was linearized by Pmel and electronically transfected into BJ5183 cells to get the recombinant adenovirus vector Ad-Runx3. The recombinant adenovirus expressing Runx3 was infected into U251 malignant glioblastoma cells. The expression of exogenous Runx3 was observed by immonoblotting and its localization was detected by immunostaining using anti-Flag tag antibody. RESULTS: The recombinant adenovirus expressing Runx3 with a Flag tag was constructed and infected into U251 glioblastoma cells. The exogenous Runx3 protein was detected only in the nuclei. CONCLUSION: The recombinant adenovirus expressing Runx3 with a Flag tag is constructed successfully, and the Runx3 protein expressed in the nuclei of infected cells. The study laid a foundation for further research of the function of Runx3 in gliocarcinogenesis.

[ABSTRACT]AIM:Toexplorethedevelopmentofhepaticsinusoidalcapillarizationintheearlystageofliverfi-brosis induced by carbon tetrachloride (CCl4) in rats.METHODS:Clean SD rats were randomly divided into normal con-trol group (group N, n=6) and liver fibrotic model group (group M, n=32).The rats in group N were intraperitoneal in-jected with saline and the rats in group M were intraperitoneal injected with CCl 4(2 mL/kg, twice a week for 4 weeks).At the end of the 3rd day and the 1st, 2nd and 4th weeks, all rats were killed and then the samples were collected .The patho-logical changes in the livers were observed by HE staining and Masson straining .The development of hepatic sinusoidal capillarization was observed by transmission electron microscopy (TEM) and immunohistochemical staining .The cell sur-face expression of vascular endothelium-associated marker CD31, collagen type Ⅳ(Col IV) and laminin (LN) was deter-mined.RESULTS:HE and Masson staining showed the formation of liver fibrosis after treatment with CCl 4 for 4 weeks. TEM showed that the fenestrate diameter of liver sinusoidal endothelial cells (LSECs) grew down, the fenestrate numbers of LSECs were decreased along with the development of liver fibrosis , and the consecutive basement membrane was formed at the end of the experiment .The expression of CD31 was significantly increased along with the development of defenestration , and the expression of Col IV and LN was significantly increased after the treatment with CCl 4 for 2 weeks and 4 weeks , re-spectively .CONCLUSION:The typical hepatic sinusoidal capillarization was detected in the early stage of liver fibrosis , and the deposition of LN in the liver sinusoidal walls was the mainly factor of formation of the consecutive basement mem -brane .