The strain, Bacillus amyloliquefaciens BF 7658 possesses the ability to produce protease and some other enzyme as well as ?-amylase. In the experiment, we have observed interference of protease with ?-amylase. If the relative ratio of activity of ?-amylase to protease was 13 : 1, 21 : 1, 27 : 1, the ?-amylase activity of culture incubated at 37℃ for 24h decreased 27.42% and 8.8% respectively. Purified ?-amylase lost the activity about 13—17% by incubation with the protease in 0.2mol/L phosphate buffer pH7.9 at 37℃ for 24h. It is confirmed that protease is one of fatters of unstability of ?-amylase.The pH and temperature affected the stability of both enzymes when cultures were heated in a 60℃ bath for 30 min. Protease activity lost about 90% and ?-amylase activity almost remained unchange.The stability of ?-amylase can be increased by adding 0.01mol/L calcium ion at pH7.0 and treating at high temperature (70℃), but protease activity is inaetivited completely. Different strains and culture conditions also affected the production ratio of ?-amylase and protease. Some strains produced high level of ?-amylase but low level of protease have been obtained.

Objective To discuss the cause, clinical features, diagnosis, treatment and prognosis of bladder and urethra leiomyoma combined with uterine leiomyoma. Methods The clinical data of 5 patients with bladder and urethra leiomyoma combined with uterine leiomyoma was analyzed retrospectively. All patients underwent surgical treatment, 3 cases were treated with enucleation of bladder leiomyoma, 1 case was treated with transurethral resection of bladder tumor (TURBT), 1 case was treated with enucleation of urethra leiomyoma,at the same time all patients were underwent hysteromyoma surgery, 1 case was treated with enucleation of partial uterine, 4 cases were treated with enucleation of hysteromyoma. Results Pathology examination showed that 4 cases were bladder leiomyoma, 1 case was urethra leiomyoma,5 cases combined with uterine leiomyoma. Patients were followed up 1-3 years,no tumor recurrence and metastasis. Conclusions Bladder and urethra leiomyoma is a kind of benign non-epithelial tumor without specific symptom,clinical rare,often merging with uterine leiomyoma, ultrasonography ,CT and cystoscopy scan could be main techniques for diagnosis mainly with surgical resection,prognosis is good. From clinical discovery bladder leiomyoma must notice patient whether at the same time merges with uterine leiomyoma.

Objective A retrospective analysis of patients with T-cell lymphoma (TCL) received autologous peripheral blood stem cell transplantation (APBSCT) was performed to evaluate the outcome of APBSCT.Methods A total of 22 patients who underwent APBSCT from September 2006 to December 2011 in Ruijin hospital were enrolled in the study,including 6 cases of lymphoblastic lymphoma and 16 of peripheral T-cell lymphoma (8 anaplastic large cell lymphoma, 4 PTCL-u, 1 subcutaneous panniculitis-like T-cell lymphoma, 2 nasal type extranodal NK/T and 1 primary cutaneous T-cell lymphoma). All patients were diagnosed based on the WHO Classification of Tumors of Hematopoietic and Lymphoid Tissues. Conditioning regimens were high-dose chemotherapies alone which include 13 cases with BEAM, 4 with ICE and 5 with CBV. The outcomes of the treatment were evaluated according to the revised International Working Group criteria.Results With a median follow-up of 13.1(1-60) months,the predicted 2-year overall survival (OS) and progression-free survival (PFS) after transplantation were (67.6±11.0) ％ and (71.1±11.1) ％,respectively.A total of 6 patients experienced disease progression and 5 patients eventually died of disease. When all these patients based on the remission status before APBSCT (CR1 vs non-CR1) and chemosensitivity (sensitive vs refractory) were further classified, the PFS rates and OS rates were 100 ％ and 91.7 ％ respectively in CR1 or chemosensitive patients which were significantly higher than patients not in CR1 (42.6 ％ ) or with chemoresistant disease (19.0 ％ ). Conclusion Remission status and chemosensitivity at the time of transplantation significantly affect the outcome of APBSCT for TCL patients, thus it can be recommend to perform APBSCT for patients either in CR1 or early stage when the disease remain sensitive to chemotherapy.

Objective Autologous hematopoietic stem cell transplantation (Auto-HSCT) has been widely used in hematological malignancies.To mobilize and harvest sufficient number of peripheral CD_(34)~+ cells is one of key issues for auto-HSCT. Peripheral CD_(34)~+ cell numeration has been used as an indicator for apheresis while we mostly rely on the peripheral WBC or MNC count. In this study, we try to evaluate the association of peripheral CD_(34)~+ count to the CD_(34)~+ cells number in the apheresis product and to find out a potential threshold. Methods From Jan 2007 to Dec 2009, a total of 57 apherosis for auto-HSCT were analysed. All patients were mobilized by cyclophophamide (CTX) plus G-CSF(5-10μg/kg) regimen. The apheresis were performed with COBE SPECTRA VERSION 6 and CD_(34)~+ count of both peripheral and apheresis products were analysed by flow cytometry. Results The median number of MNC in apheresis products was 4.6(0.3-10.5)×10~8/kg with median CD_(34)~+ cells at 2.4(0.16-34.9)×10~6/kg. The peripheral CD_(34)~+ count was the only parameter associated with the MNC and CD_(34)~+ cell numbers in the apheresis products while the WBC number was irrelevant to the results of apheresis. Our data showed that when the peripheral CD_(34)~+ count reach 15/μl, the efficacy of a single apheresis significantly improved with 81 % and 60 % reached 1 and 2×10~6 CD_(34)~+ cells/kg respectively and the total number of MNC and CD_(34)~+ cells were significantly superior to apheresis with peripheral CD_(34)~+ cells <15/μl, thus indicated that CD_(34)~+ ≥15 /μl can be used as the threshold for apheresis. Furthermore, the ROC analysis demonstrated that CD_(34)~+ cells ≥25(26.5-28.6) /μl is the best indicator level for a successful single apheresis. Conclusion Our study clearly showed that peripheral CD_(34)~+ cell count is a key indicator of apherosis. CD_(34)~+ cells at 15/μl can be used as the threshold to start apheresis in the clinical setting.

Objective:To investigate the inhibitory effect of small interfering RNA-Yes-associated protein 1 (siRNA-YAP1) on epithelial-mesenchymal transition (EMT) in human lens epithelial cells (LECs) induced by transforming growth factor-β 2 (TGF-β 2). Methods:Human LECs line (HLEB-3) was cultured and divided into normal control group and TGF-β 2 induced group.The cells in the normal control group were treated with serum-free low-glucose medium for 24 hours, and the cells in the TGF-β 2 induced group were treated with additional 10 ng/ml TGF-β 2 for 24 hours.The cultured HLEB-3 cells were divided into siRNA empty vector group, siRNA-YAP1 transfection group, siRNA empty vector+ TGF-β 2 group and siRNA-YAP1+ TGF-β 2 group, and the cells were transfected with plasmid including siRNA empty vector or siRNA-YAP1 sequence according to grouping.The relative expression levels of YAP1 mRNA and protein in various groups were detected and compared by quantitative real-time polymerase chain reaction (PCR), immunofluorescence and Western blot assay, respectively.The relative expression levels of EMT marker proteins (E-cadherin and Vimentin proteins) in various groups were detected by immunofluorescence and Western blot assay. Results:Compared with the normal control group, the expression level of E-cadherin protein was decreased (1.180±0.118 vs.0.830±0.104) and the Vimentin protein was increased (0.797±0.110 vs.1.240±0.110) in the TGF-β 2 induced group, with significant differences between the two groups ( t=3.857, P=0.018; t=-4.933, P=0.008).The relative expression levels of YAP1 mRNA and protein in the TGF-β 2 induced group were significantly increased in comparison with the normal control group (2.200±0.193 vs.1.136±0.123; 1.203±0.121 vs.0.967±0.025), with significant differences between the two groups ( t=-9.288, P<0.01; t=-3.329, P=0.029).Compared with the siRNA empty vector group, the expression levels of YAP1 mRNA and protein in the siRNA-YAP1 transfection group were significantly reduced (both at P<0.01).Compared with the siRNA empty vector+ TGF-β 2 group, the relative expression level of E-cadherin protein was significantly enhanced and the expression level of Vimentin protein was significantly reduced in the siRNA-YAP1+ TGF-β 2 group (both at P<0.01). Conclusions:YAP1 participates in the TGF-β 2 induced EMT in human LECs, and siRNA-YAP1 can suppress the EMT process.

Objective To investigate the clinical characteristics and survival analysis of T-cell lymphoma with hemophagocytic syndrome (HPS).Methods The clinical features,laboratory tests and survival of 30 patients of T-cell lymphoma with HPS were analyzed comparing with 50 random patients of T-cell lymphoma without HPS (control group),and the data were obtained from department of hematology,Ruijin Hospital,Shanghai Jiaotong University School of Medicine from January 2006 to December 2011.Results 30 cases of T-cell lymphoma with HPS manifested for high fever [100.00 ％ (30/30)],splenomegaly [96.67 ％ (29/30)],more than two-line reduction in blood cells [93.33 ％ (28/30)],bone marrow with bloodthirsty phenomenon [86.67 ％ (26/30)],LDH increased [100.00 ％ (30/30)],hypertriglyceridemia [46.67 ％ (14/30)],low fibrin hyperlipidemia [60.00 ％ (18/30)],ferritin increased [93.33 ％ (28/30)],liver damaged [86.67 ％ (26/30)] from patient group,which were higher than those of control group,the differences were statistically significant (x2 =23.11,22.50,36.05,64.20,21.82,5.31,16.54,26.82,46.43,all P ＜ 0.05).But the incidence of superficial lymph nodes enlargement was not high,only 33.33 ％,which made the diagnosis of primary disease difficult.Combined chemotherapy of CHOP or etoposide plus dexamethasone could make the symptoms and laboratory indicators improved,but the median survival time was only 20 days.Compared with the control group,there were significant differences on the survival curves (x2 =35.05,P ＜ 0.0001).Conclusion In T-cell lymphoma with HPS,the clinical manifestations are complex,usually with multi-organ dysfunction,aggressive course,and poor prognosis.The pathogenesis and treatment of HPS should be further studied.

Objective To evaluate adjuvant chemoradiotherapy on gallbladder carcinoma after RO resection.Methods A total of 54 patients undergoing curative resection (R0) for gallbladder carcinoma between January 1991 and December 2006 were recruited for evaluation on the effect of postoperative adjuvant chemoradiotherapy,with the exception of 5 TINOM0 patients.Among the remaining 49 patients,11 patients received radiotherapy,and 21 cases had undergone adjuvant chemotherapy.Results The overall 1,3 and 5-year survival rates of these 54 patients were 81.1％,54.4％ and 48.4％,respectively.The overall 5-year survival rates of T1,T2,T3 and T4 were 100％,92.3％,27.8％ and 12.5％ ( x2 =25.307,P ＜0.001 ),respectively.The median survival for those patients receiving adjuvant radiotherapy was 12.8 months compared to 28 months of median survival for those treated without adjuvant radiation therapy (x2 =3.942,P =0.047 ).Adjuvant radiotherapy significantly prolonged overall median survival from 16 months to 95 months ( x2 =5.387,P =0.02 ) in the subgroup of patients with T3/T4 stage.Adjuvant radiotherapy for patients with positive lymph nodes improved overall median survival from 12 to 19 months (x2 =3.959,P =0.047 ).In addition,radiotherapy prolonged overall median survival from 12 moths to 128 months in patients with G3/G4 disease ( x2 =7.401,P =0.007).The median survival was not significantly different among patients with or without adjuvant chemotherapy (x2=0.086,P=0.770 ).Conclusions Adjuvant radiotherapy is recommended for lymph node positive,T3/T4,or G3/G4 gallbladder carcinoma patients following R0 resection.

Objective To evaluate the efficacy and safety of ureteroscopic endo-incision and drainage in the treatment of renal cysts.Methods Thirty cases(19 females and 11 males)of renal cysts(1 1 parapelvie cysts,15 simple cysts.and 4 multiple cysts)were treated with ureteroscopic endo-incision and drainage.The renal cysts were located in renal pelvis,and opened and decompressed by electrotme with ureteroscope.Double J stent was placed afterwards.Urinary and blood biochemistry were tested post-operatively.Results All the operations were successfully completed with no severe complication.The cyst managing time ranged from 15 tO 45 min.Urinary biochemistry(urinary protein and glucose)turned normal 1 2 days after the surgery.Patients were followed up for 3 to 9 months.Renal cysts disappeared in 24 cases,diminished in 4 cases,and recurred in 2 cases.Conclusion Application of ureteroscopic technique in the treatment of renal cyst is safe,effective and minimally invasive.

Objective To investigate the effects of chronic hepatitis B virus (HBV) infection on human hepatic cytochrome P450 2C9 (CYP2C9).Methods Liver tissue samples and blood samples were obtained from 10 patients with chronic HBV infeetion and 10 healthy controls.CYP2C9 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.The activity of CYP2C9 was detected utilizing high performance liquid chromatography (HPLC).The expressions of CYP2C9 mRNA and protein were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blotting.The data were analyzed by t test.Results All the liver samples showed CYP2C9 wild-type (*1*1),while CYP2C9 (*2) and CYP2C9 (*3) were not detected.The maximum velocity (Vmax) of CYP2C9 in patients chronic HBV infection and healthy controls were (263.5±66.4) μmol/L and(284.6±85.9) μmol/L,respectively (t=0.614,P=0.5471).The expression of CYP2C9 mRNA in patients with chronic HBV infection (0.39±0.28) was significantly lower than that of healthy controls (0.65±0.13) (t=2.628,P=0.0171).Accordingly,the protein expression in patients with chronic HBV infection (0.26±0.13) was lower than that of healthy controls (0.60±0.19) (t=4.688,P=0.000 2).Conclusion The expressions of CYP2C9 mRNA and protein are decreased in chronic HBV infection which may down-regulate the enzyme activity.