The primary structure of TNK-tissue plasminogen activator (TNK-tPA) was characterized using liquid chromatography-mass spectrometry (LC-MS). Firstly, the molecular mass of deglycosylated protein was measured. Then peptide mass mapping and MS/MS of the reduced, alkylated and trypsin-digested sample were tested and analyzed so as to verify its amino acid sequence and identify post-translational modifications. Results show that the amino acid sequence was consistent with designed structure; about 5% of M207 was oxidized; T61 was fucosylated with -80% occupancy; N103, N448 and N184 (-15% occupancy) were glycosylated with complex-type oligosaccharides. LC-MS coupled with proper sample pretreatment is approved to be a rapid and powerful approach to characterize the primary structure of TNK-tPA.

ObjectiveTo compare the clinical effectiveness and adverse effects following low doses versus traditional doses of amphotericin B liposome (L-AmB) in the treatment of patients with invasive pulmonary fungal infections (IPFI) after renal transplantation.MethodsA total of 26 postrenal transplantation patients with IPFI between Jan. 2005 and Mar. 2011in Zhujiang hospital received L-AmB treatment identified low doses group (0.2-0.5 mg·kg-1·d-1,n =19) or traditional doses group (1-5 mg· kg-1,d-1,n =7) were reviewed.ResultsThe treatment duration in low doses group and traditional doses group was 20.3 +12.7 and19.3 ±13.2 days respectively (P＞0.05).The effective rate in low doses group and traditional doses group was 84.2％ and 57.1％ respectively (P＞0.05).The overall dosage was significantly less in the low doses group (414.7 ± 241.7 mg) than in the traditional doses group (1158.8 ± 928.0 mg) (P＜0.05).The incidence of adverse effect was significantly lower in the low doses group than in the traditional doses group (21.1％ vs.85.7％,P＜0.05).ConclusionThe effectiveness of low doses of L-AmB protocol in the treatment of IPFI postrenal transplantation patients was similar to that of traditional doses of L-AmB protocol,but the incidence of adverse effects in low doses of L-AmB protocol was significantly lower.

The angiotensinogen(AGT) expression and angiotensin Ⅱ (AngⅡ ) secretion levels in cultured SD rat mesangial cells were determined. High glucose up-regulated AGT mRNA(0. 29±0.07 vs 0. 20±0. 05,P< 0.05)and protein(0.66±0.23 vs 0.37±0. 15,P<0.05) expression and Ang Ⅱ secretion [(9.85±2.08 vs 7.50± 1. 51) pg/ml,P<0. 05]levels, which were down-regulated by pyrrolidine dithiocarbamate( PDTC) treatment via inhibiting NF-κB activity.

The relationship between C936T polymorphism at 3'-untranslated region of vascular endothelial growth factor (VEGF) gene and diabetic nephropathy (DN) was analysed in 194 type 2 diabetic patients. The frequencies of genotype CC and allele C were significantly higher in DN group than those in non-DN group and control group. Allele C and genotype CC of VEGF may be a genetic marker susceptible to DN.

BACKGROUND: Recent studies have suggested that conversion from cyclosporine A (CsA) to tacrolimus (FK 506)-based regimen can improve renal allograft function and survival rate. But little is known about whether the conversion from CsA to tacrolimus(FK 506) plus mycophenolate mofetil (MMF)-based regimen exhibits the same or similar clinical efficacy. OBJECTIVE: To investigate the clinical efficacy and safety of converting CsA to FK506 plus MMF in treatment of different types of chronic allograft nephropathy (CAN). DESIGN, TIME AND SETTING: An observational and controlled trial was performed at the Center for Organ Transplantation, Zhujiang Hospital, Southern Medical University from January 2005 to October 2007. PARTICIPANTS: Fifteen-nine enrolled patients received CsA-based regimen after renal allografting. Following pathological confirm and typing, all patients were assigned to two groups: CAN with chronic rejection (CR, n = 31) and CAN without chronic rejection (non-CR, n = 28). FK 56 was purchased from Fujisawa Pharmaceutical Company, Ltd., Japan. MMF was sourced from Shanghai Roche Pharmaceutical Co., Ltd., China. METHODS: When patients were diagnosed CAN, the CsA regimen was conversed to FK506 plus MMF regimen. FK506 initiated at a dose of 0.08 mg/kg per day and then was adjusted to achieve steady-state whole blood trough levels of approximately 5-8 μg/L. MMF was used at a fixed dosage, 1.0 g/d, twice a day, only if relative adverse events occurred. All patients were followed up at least 6 months. MAIN OUTCOME MEASURES: Serum creatinine(Scr), total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), 24-h proteinuria, glomerular filtration rate (GFR), and complications. RESULTS: All initial 59 patients were included in the final analysis. At 6 months after regimen conversion, the levels of Scr, TC, TG, LDL, and 24-hour proteinuria were significantly reduced in non-CR, in particular CR, groups, compared with prior to conversion (P< 0.05). GFR was markedly increased in both the CR and non-CR groups (P< 0.05). In the CR group, 20 patients obtained improved results, 7 got stable results, and 4 showed ineffective results. The effective rate of regimen conversion was 64.5% and 32.1% in the CR and non-CR groups, respectively, and significant difference existed between the two groups (P < 0.05). Compared with prior to conversion, the incidence of hypertension and hyperlipemia was significantly decreased after regimen conversion (P< 0.05). There was no significant difference in diabetes mellitus, opportunistic infection, and malignancy between prior to and after regimen conversion. CONCLUSION: FK506 plus MMF-based regimen can markedly improve the function of renal graft of CAN, in particular CR, patients.

Objective To explore the changes serum S-100? in children with acute carbon monoxide poisoning and its clinical significance.Methods The levels of serum S-100? of 28 children with acute carbon monoxide poisoning and those of 20 healthy children were mea-sured by enzyme-linked immunosorbent assay.Results The serum S-100? levels of the study group and control group were(0.517?0.346)and(0.037?0.014)?g/L respectively,there was significant difference between two groups(t=6.197 P

ObjectiveTo investigate the risk factors for contrast-induced nephropathy (CIN) after endovascular therapy in patients with peripheral arterial diseases and to evaluate the conformance of serum cystatin C (Cys C) and serum creatinine in diagnosis of CIN. Methods In this prospective,single center study,in-hospital patients with peripheral arterial diseases undergoing non-emergency endovascular therapy from July 2010 to April 2011 in our hospital were enrolled.CIN was defined asScr increase ≥25％ after angiography.General clinical characteristics and blood biochemical parameters were compared between the non-CIN and CIN groups.Logistic regression analysis was performed to determine risk factors.Changes compared to baseline level in serum creatinine and Cys C at predefined time-points were evaluated.Results A total of 367 patients were enrolled in the study.The proportions of patients with diabetes mellims and treatment with diuretics before angiography,contrast-media dosage were significantly higher in the CIN group than those in non-CIN group (P＜0.05,＜0.01,＜0.01).Logistic regression analysis indicated that diabetes mellitus,contrast-media dosage were risk factors for CIN.Several serum Cys C increase criteria at 24 hours after contrast media exposure all had low sensitivity for predicting a Scr increase ≥25％.Only small overlapped regions were found in Venn diagram between several increasing criteria according to serum Cys C and serum creatinine criterion. Conclusions Diabetes mellitus,contrast-media dosage are independent risk factors for CIN.The results of several increasing criteria according to serum Cys C in evaluating contrast-induced AKI are not coincident well with that of serum creatinine criterion.

To establish a detection method of oncolytic adenovirus/p53 and standard of quality control, human telomerase reverse transcriptase (hTERT) promoter, CMV fusion promoter containing hypoxia reaction element (HRE) and p53 gene were identified by vector DNA restriction enzyme digestion and PCR analysis. The result conformed that all modified regions were in consistent with theoretical ones. Particle number was 2.0 x 10(11) mL(-1) determined by UV (A260). Infectious titer was 5.0 x 10(10) IU mL(-1) analyzed by TCID50. In vitro p53 gene expression in human lung cancer cell H1299 was determined by ELISA, and A450 ratio of nucleoprotein in virus infection group to control group was 5.2. Antitumor potency was evaluated by cytotoxicity assay using human lung cancer cell A549, and the MOI(IC50) of this gene therapy preparation was 1.0. The tumor cells targeted replication ability of recombinant virus was determined by TCID50 titer ratio of filial generation virus between human lung cancer cell A549 and human diploid epidermal fibrolast BJ cells after infected by virus with same MOI. TCID50 titer ratio of tumor cell infection group to normal cell infection control group was 398. The IE-HPLC purity of virus was 99.5%. There was less than 1 copy of wild type adenovirus within 1 x 10(7) VP recombinant virus. Other quality control items were complied with corresponding requirements in the guidance for human somatic cell therapy and gene therapy and Chinese pharmacopeia volume III. The detection method of oncolytic adenovirus/p53 was successfully established for quality control standard. The study also provided reference for quality control of other oncolytic viral vector products.

Background and purpose:Hypermethylated in cancer 1 (HIC1) is silenced in multiple cancer cells and tissues by DNA methylation of epigenetic modification, which may modulate the initiation and progression of tumors. However, there are few reports about this phenomenon in prostate cancer. This study aimed to investigate the status of HIC1 promoter methylation in prostate cancer using methylation methods.Methods:Methylation-specific polymerase chain reaction (MSP) and bisulfate sequencing PCR (BSP) were used to detect the methylation status ofHIC1 promoter in prostate cancer cell lines PC3 and C4-2B, prostate normal cell line PrEC, primary Chinese PCa tissues and the respective healthy control cases.HIC1 expression level was respectively determined by reverse transcription-PCR (RT-PCR) and Western blot assays in PC3, C4-2B and PrEC cells treated with 5-Aza-CdR.Results:We found that the percentages of HIC1 promoter methylation were 78.23%, 72.15% and 10.63% in PC3, C4-2B and PrEC cells by MSP analyses. Moreover, the levels of methylatedHIC1 promoter in 36 primary Chinese PCa tissues compared with the respective healthy control cases were 80.30%vs 31.56%. Expressions ofHIC1 mRNA and protein level were restored in PC3 and C4-2B cells after 5-Aza-CdR treatment.Conclusion:These findings demonstrate thatHIC1 promoter region is hypermethylated in prostate cancer, which results in silence or downregulation ofHIC1. The status ofHIC1 methylation can be a valuable marker in the early stage of prostate cancer and a potential therapeutic target.

Objective To summarize the puncture indications and the pathological type features of renal allografts biopsies in our center for evaluating its safety and diagnostic value .Methods The data of 773 percutaneous renal allograft biopsies in 629 kid‐ney transplants in the Pearl River Hospital of Southern Medical University from January 2005 to June 2014 were retrospectively an‐alyzed .Results The success rate of renal biopsy was 100% ,9 cases(1 .2% ) were complicated postoperative perirenal small hemato‐ma ,33 cases(4 .3% ) with gross hematuria and 1 case(0 .13% )with abdominal pain .Among the indications of 773 biopsies ,protein urine occured 205 cases(26 .5% ) of patient ,blood Cr increased in 187 cases(24 .2% )of patients ,protein urine simultaneously com‐plicating blood Cr increased ,in 313 cases of patients ,53 cases(6 .9% )had postoperative oliguria urinary ,and 15 cases(1 .9% )were get procedural biopsy .In the pathological types ,21 cases(2 .7% ) were normal ,179 cases (23 .2% ) were acute T cell‐mediated rejec‐tion after transplantation ,51 cases (6 .6% )were acute antibody‐mediated rejection ,205 cases (26 .5% ) were chronic T cell‐mediated rejection and 43 cases(5 .6% ) were chronic antibody‐mediated rejection;41 cases(5 .3% ) were drug toxicity ,29 cases(3 .7% ) were acute tubular necrosis(ATN) ,11 cases(1 .4% ) were relapsed or new nephropathy ;9 cases(1 .2% )were HBV related renal disease;39 cases (5 .0% ) were critical lesion and 145 cases(18 .8% )were others .Conclusion Rrenal allograft biopsy is safe ,it is important to the etiological diagnosis of renal disease after renal transplant ,which can guide the clinical treatment and improve the long term survival of renal graft and should be routinely carried out in clinic .