Objective To present 3 cases of MFH of the urinary system. Methods Two cases of renal MFH and one case of bladder MFH were retrospectively analysed. Results The diagnosis was established on pathological examination and immunohistochemistry. Two patients died 4～7 months affer operation , while the other patient has been well for 30 months. Conclusions It is difficult to differentiate it from primary renal and bladder neoplasm.Immunohistochemistry is helpful to distinguish it from carcinoma,renal sarcoma and renal parental cell carcinoma.

Objective To discuss the outcome of repair of limbs bone defect by using mineralized collagen graft.Methods The clinical data of 35 cases who diagnosed the existence of bone defect from January 2013 to December 2013 in our department were retrospectively analyzed.After operation,Lane Sandhu X-ray score standard was evaluated.Results All patients were followed up for 12 months.Postoperative 1 month,Lane Sandhu X-ray score was 3 points in 20 cases and 15 cases with 2 pointss;postoperative 6 months,Lane Sandhu X-ray score was 9 points in 21 cases,10 cases with 8 points and 4 cases with 7 points.Postoperative 12 months,Lane Sandhu X-ray score was 12 points in 28 cases,7 cases with 11 points.The scores of postoperative 6 months were better than the scores of postoperative 1 month [(2.60±0.49)points vs.(8.49±0.12)points,t=107.860,P=0.000].There was significant difference between the scores of postoperative 12 months and scores of postoperative 6 months [(8.49±0.12)points vs.(11.8±0.06)points,t=41.630,P=0.000].Conclusion MC is used as an effective bone substitute material,and its clinical effect is good.

ObjectiveTo evaluate radio-frequency hemostasis in hepatectomy.MethodsFrom January 2009 to February 2011,the clinical data of 60 patients undergoing curative liver resection were divided into two groups using radio-frequency hemostasis (RFH) and clamp crushing method (CCM) respectively,RFH group (30 cases) and CCM group (30 cases).There was no difference between the 2 groups regarding the age,sex.hepatic function and tumor size.Data regarding the intra-operative and postoperative courses of the patients were analyzed.ResultsNo damage of hepatic vein occured in RFH group.Hepatic veins rupture occurred in 5 cases and massive bleeding occurred in 3 cases in CCM group.lntra-operative blood loss was significantly less in FRH group [ (219 ±62) ml] than in CCM group [ (416 ±96) ml ] (P ＜ 0.05 ).The postoperative drainage volume in RFH group was significantly less than that in CCM group on the third postoperative day.The serum ALT and T-BIL in RFH group was significantly lower than that in CCM group on postoperative day 1 and day 7 ( separately t =5.987,16.803,22.264,8.386,8.255,all P ＜0.05 ).Postoperative hepatic function in RFH group was significantly better than that in CCM group.ConclusionsThe use of radio-frequency hemostasis in hepatectomy is less traumatic,of less bleeding,faster recovery than clamp crashing method.

Objective:To investigate specific cellular immune response to the HPV16E7 prophylactic vaccine in mice.Methods:BALB/c mice were randomly divided into 3 groups:experimental group (treated with pcDNA3.1-HPV16E7),control group Ⅰ(treated with pcDNA3.1) and control group Ⅱ(treated with N.S.).Mice were injected (i.m.) pcDNA3.1-HPV16E7,pcDNA3.1 and N.S. one time per week,respectively.After three immunization,the blood samples from eye sockets and the supernatant cultured of spleen cells were taken for measurement IFN-? and the number of CD4 +?CD8 +T-lymphocyte by ELISA and FACS assay.Antigen-specific splenocyte proliferation assay in vitro was detected by MTT method.Results:The splenocytes actively proliferated,the number of CD4 +T lymphocyte and the quantitation of IFN-? in spleen and serum in the experimental group were significantly higher than the control group Ⅰ and Ⅱ.Conclusion:The pcDNA3.1-HPV16E7 DNA vaccine can induce specific cellular immune response in BALB/c mice.

Objective 　To establish a new practical method to assess the cerebral blood flow autoregulation. Methods　We assessed the flow velociey of middle cerebral artery with transcranial Doppler and recorded invasively the blood presure simultaneonsly. Then on the basis of critical closing pressure (CCP), the lower limit of cerebral blood flow autoregulation and the blood flow resistance of arterioles were calculated.The data compared with the results generated by routine method. Results　The lower limit of autoregulation working out by CCP was 70.88±24.05 mmHg, which was similar to the result measured by routine method. The lower limit of autoregulation and the arteriole resistance in RHR were significantly higher than those of normal controls, and highly relate to arterial blood pressure significantly, especially pulse pressure. Conclusions　The physiology and pathology of cerebral blood flow can be evaluated conveniently and accurately by assessment of the lower limit of autoregulation and arterioles resistance with CCP.

AIM:To study the effect of hypertensive arteriosclerosis on cerebral blood flow autoregulation (CBFA), and to introduce a new method to measure the lower limit. METHODS:The blood velocities and blood pressure was recorded simultaneously and the curves of CBFA were analyzed and classified into classical and non-classical pattern. The lower limit were determined by clinical closing pressure (CCP) and the curve CBFA. RESULTS:When the blood pressure was decreasing, the classical and non-classical pattern of the cerebral blood flow autoregulation were 25% and 75% respectively in normal SD rats, while they were 40.55% and 54.45% respectively in renovascular hypertensive rats (RHR). However, when the blood pressure was elevating, the classical and non-classical pattern were 76.47% and 23.53% respectively in SD rats, while they were all classical in RHR. Furthermore, in SD and RHR ras, the lower limits measured by CCP were well in accordance with that measured by CBFA. CONCLUSION:Hypertensive arteriosclerosis could influence the limits and the patterns of cerebral blood flow autoregulation. The lower limit of CBFA can be measured and analyzed by CCP.

Objective To investigate mechanism of netrin-1 regulating invasion of extra villous trophoblasts. Methods RT-PCR was used to detect six receptors expression including UNC5A, UNC5B, UNC5C, UNC5D, DCC and neogenin in extra villous trophoblast cell line TEV-1. The TEV-1 cells were cultured and devided into seven groups according to the concentration of netrin-1 adding into the medium, which include 10 μg/L, 50 μg/L, 100 μg/L, 500 μg/L, 1000 μg/L, 5000 μg/L and the control(the concentration of netrin-1 was 0 μg/L) groups. The proliferation and invasion of TEV-1 induced by netrin-1 were determined by CCK-8 assay and transwell invasion assay respectively. Results (1) Only neogenin and UNC5B were found to be expressed on TEV-1 by RT-PCR method. (2) In CCK-8 proliferation assay, after 72 hours culture, the proliferation of TEV-1 were 1.55 ±0.29 in 10 μg/L, 1.72±0. 31 in 50 μg/L, 2.15 ±0.35 in 100 μg/L, 1.42 ±0. 25 in 500 μg/L, 1.50±0. 27 in 1000 μg/L, and 1.38±0.23 in 5000 μg/L group, which were all higher than 1.00 ± 0.16 in control group significantly ( P<0.05 ). (3) In matfigel invasion assay, after 6 hours culture, the number of the trans-membrane cells in various netrin-1 group, including 41 ±4 in 10 μg/L, 47 ±5 in 50 μg/L, 55±6 in 100 μg/L, 44 3=5 in 500 μg/L, 43±5 in 1000 μg/L and 42 ±5 in 5000 μg/L group, were all higher than 30 ±4 in control group with statistical significance( P<0.05 ). (4) The fold changes of neogenin were 1.50 ± 0.16 in 10 μg/L, 1.83 ± 0.19 in 50 μg/L, 2.24 ± 0.25 in 100 μg/L, 2.12 ±0.24 in 500 μg/L, 2.12±0.23 in 1000 μg/L and 2.13 ± O. 23 in 5000 μg/L group, which were all higher than 1.00 ±0.11 in control group significantly( P <0.05 ). There were significant difference between group 10 μg/L and 50 μg/L, group 50 μg/L and 100 μg/L (P < 0.05). There were no significant difference between group 100 μg/L and 500 μg/L, group 1000 μg/L and 5000 μg/L (P>0.05). (5) The fold changes of UNC5 B 1.09 ± 0.11 in 10 μg/L, 1.47±0.14 in 50 μg/ L, 1.61 ±0.16 in 100 μg/L, 1.85±0.19 in 500 μg/L, 2.21±0.21 in 1000 μg/L and 2.42±0.23 in 5000 μg/L group, were all higher significantly when compared with 1.00 ± 0.07 in control group (P<0.05 ). There were significant difference between all groups ( P<0.05). Conclusion Netfin-1 can promote the potential of proliferation and invasion of extravillous trophohlasts in vitro through its receptors including neogenin and UNC5 B.

Objective To construct the eukaryotic expression plasmids of human Fas and TNFR1 gene(pcDNA3.0-hFas and pcDNA3.0-hTNFR1)and microRNA(miRNA)expression plasmid of hFas and hTNFR1 named p-hFasmiRNA and p-hTNFR1miRNA,and to investigate their inhibitory effects in vitro.Methods The eukaryotic expression plasmids of human Fas and TNFR1 gene were constructed(pcDNA3.0-hFas and pcDNA3.0-hTNFR1)and have been shown successfully to express hFas and hTNFR1 protein.miRNA expression plasmids of hFas and hTNFR1 named p-hFasmiRNA and p-hTNFR1miRNA complimentary to the sequence responsible for hFas and hTNFR1 respective were constructed,meanwhile irrelevant miRNA plasmid was used as a control.By respective co-transfection of p-hFasmiRNA and pcDNA3.0-hFas,p-hTNFR1 miRNA and pcDNA3.0-hTNFR1 expression construct into 293T cells,the inhibition of hFas and hTNFR1 expression was analyzed by real-time PCR and Western blot.Results The experiments showed the significant inhibitory effect of p-hFasmiRNA on hFas and p-hTNFR1miRNA on hTNFR1 expression at 48 h post-transfection both at RNA level and protein level as well in 293T cell lines with the inhibitory efficiency being as high as 87% for hFas and 80% for hTNFR1,respectively.Conclusion The p-hFasmiRNA and p-hTNFR1miRNA were constructed successfully,and it was verified that they could specifically inhibit the hFas and hTNFR1 expression at the cellular level.

Previous study on TNFRl-mediated hepatocyte apoptosis has been implicated in the development of fulminant viral hepatitis.To interfere with the potentially effective target,plasmid named p-mTNFR1shRNA complimentary to the sequence responsible for mTNFR1 was also constructed and further confirmed by sequence analysis.To investigate the effect of mTNFR1shRNA plasmid on mTNFR1 expression in vivo and the disease progress in MHV-3 induced fulminant hepatitis mice model.By hydrodynamic injection of mTNFRlshRNA plasmid,the survival rate of mice,hepatic pathological change were examined and compared between mice with/without mTNFR1 shRNA plasmid intervention.The expression of mTNFR1 was detected by Real-time PCR,immunohistochemistry staining.The mTNFR1 shRNA plasmid significantly reduced mTNFR1 expression in vivo,markedly ameliorates inflammatory infiltration,prolonged the survival time period and elevated the survival rate from 0 up to 13.3% in Balb/cJ mice with MHV-3 induced fulminant hepatitis.This study was designed to explore the opportunity of RNA interference technique in inhibiting TNFR1 expression,which has been reported to be involved in the development of a variety of diseases including fulminant viral hepatitis and severe chronic hepatitis B.

Objective To evaluate the safety and efficacy of bipolar plasmakinetic system in exfoliative tran-surethral resection of bladder tumors .Methods Clinical data of 72 patients with non muscle invasive bladder cancer (NMIBC) were retrospectively analyzed.Transurethral bipolar plasmakinetic system was used ,30°viewer,F27 outer sheath was pushed off and bladder tumor was cut .When the bladder filling state ,pushed off bladder mucosa distance from tumor basal 2cm,then electricity cut the exfoliative bladder tumor .When bladder half filling state electricity cut the base of the bladder muscle layer of bladder tumor .Results This group of 72 cases were successfully completed surgery,surgery time 37~93 min,without intraoperative bladder perforation ,slight obturator nerve reflex in 5 cases. The keeping intact pathologic specimens was good for pathological staging .Conclusion Exfoliative transurethral resection of bladder tumors with bipolar plasma is a safe , practical and effective way of operation , which can avoid severe obturator reflex occurred in the operation , and greatly reduce the occurrence of bladder perforation , without TURS,surgical removal of the pathological specimens is specification .