[Clinical value of ATP determination in CD4+ cells of patients with cytomegaloviral pneumonia after kidney transplantation].

Objective: To explore the clinical value of determination of ATP levels in CD4(+) cells of patients with cytomegaloviral pneumonia after kidney transplantation.

Methods: Twenty-eight patients with cytomegaloviral pneumonia following kidney transplantation and 30 healthy volunteers were enrolled in this study. ATP-bioluminescence assay (ATP-CVA) was used to assess the immune response of CD4(+) cells to phytohemagglutinin (PHA) stimulation in the normal volunteers and the recipients (before and at 1, 2, and 4 weeks after renal transplantation, before and at 2 and 4 week after the treatment).

[Comparison of the surgical approaches for nephrectomy in living related donors].

Objective: To compare the clinical effects and graft outcomes of 4 surgical approaches for nephrectomy in living related kidney donors.

Methods: Between June, 2004 and June, 2007, 119 living related kidney donors underwent nephrectomy via different surgical approaches, and their clinical data were retrospectively analyzed. Of these donors, 22 received retroperitoneal open nephrectomy, 21 had retroperitoneoscopic nephrectomy, 13 had hand-assisted laparoscopic nephrectomy, and 63 underwent transperitoneal open nephrectomy.

[Etiologic investigation of chronic granulomatous inflammation of hand by polymerase chain reaction and DNA sequencing].

Objective: To explore the causative pathogens in littoral hand infections which exhibited chronic granulomatous inflammation, the relationship between chronic granulomatous inflammation and mycobacteria and to discuss the prospects of PCR in clinical application for diagnosis of granulomatous inflammation.

Method: With 16S-rDNA as the target sequence, Nest-PCR was used to detect mycobacteria directly from 37 cases of chronic granulomatous inflammations, and identified them by gene sequencing.

Results: Twenty-four of 37 cases were positive for mycobacteria by Nest-PCR, in which 17 were M.

[Half-dose Zenapax for acute rejection prevention after renal transplantation].

Objective: To investigate the efficacy and safety of half-dose Zenapax for prevention of acute rejection after renal transplantation.

Methods: According to the immunosuppressive regimen and renal function after transplantation, patients were divided into 4 groups, namely groups A, B, C, and D of 90, 73, 11 and 13 patients, respectively. Blood creatinine measured 1 week after operation was <176.

[Management of urinary obstruction following renal transplantation: report of 16 cases].

Objective: To explore the diagnosis and treatment of urinary obstruction involving the transplanted kidney.

Methods: A retrospective analysis was performed in 16 cases of urinary obstruction involving the transplanted kidney, including 5 cases of ureteral calculi, 6 vesicoureteral anastomotic stricture, 2 pyeloureteral junction stricture after transplantation, 1 ureter necrosis due to graft rejection, and 2 infection surrounding the renal graft and ureter end necrosis.

Results: Only one patient had the renal graft removed due to massive hemorrhage in an open surgery for correction of urinary obstruction, and the renal function of the graft was preserved in all the other cases after endoscopic or open surgeries.

[Expression of IgG receptor on human umbilical vein endothelial cells stimulated with TNF-alpha and IFN-gamma].

Aim: To detect the expression of IgG receptors (FcgammaR) on cytokine-stimulated human umbilical vein endothelial cells (HUVECs).

Methods: By using ELISA, immunocytochemical staining, immunofluorescent staining and RT-PCR, the expression and subtypes of FcgammaR were detected.

Results: Non-stimulated HUVECs expressed very low level of FcgammaRIIa.

[Differential analysis of tyrosine-phosphorylated proteins in human hepatocellular carcinoma cell lines with different metastasis potentials].

Objectives: To compare expressions of tyrosine-phosphorylated proteins in different hepatocellular carcinoma cell lines with different metastasis potential and to screen key molecules associated with HCC metastasis and recurrence.

Methods: Using two-dimensional electrophoresis, Western blotting and MALDI-TOF-MS/MS, we analyzed tyrosine-phosphorylated protein profiles of Hep3B, MHCC97L and MHCC97H, HCC cell lines with different metastasis potentials.

Results: 10 spots were detected in Hep3B, 19 in MHCC97L and 17 in MHCC97H.

Identification of a novel nucleus protein involved in the regulation of urokinase in 95D cells.

The urokinase-type plasminogen activator (uPA) plays an important role in cellular invasion. By using the downstream part of a 74 bp DNA region called the cooperation mediator (COM) of the uPA promoter as a bait sequence in the yeast one-hybrid screen, a gene called PBK1 was previously cloned from the cDNA library of the 95D lung cancer cell strain. In this study, the intracellular distribution of PBK1 was studied by using the transient transfection of pEGFP-C3-PBK1, and PBK1 was found to be localized in the nucleus.

Interaction between plasminogen activator inhibitor type-2 and pre-mRNA processing factor 8.

The plasminogen activator inhibitor type-2 (PAI-2) dependent apoptosis protection is due to the 33 amino acids fragment located between helix C and D of PAI-2, this fragment may interact with some unknown intracellular proteins. In this study we used the fragment between helix C and D of PAI-2 as a bait to perform a yeast two-hybrid screen using a cDNA library constructed with HeLa cells during apoptosis, and retrieved a clone encoding 94 amino acid residues of C-terminus of pre-mRNA processing factor 8 (PRPF8). Co-immunoprecipitation experiments confirmed that PAI-2 could interact with PRPF8 in vivo.

Antisense Tiam1 down-regulates the invasiveness of 95D cells in vitro.

As a specific guanine nucleotide exchange factor of Rac1, Tiam1 (T-lymphoma invasion and metastasis inducing protein 1) is involved in a number of cellular events, such as cytoskeleton reorganization, cell adhesion, and cell migration. Since Tiam1 was implicated in the invasion and metastasis of T-lymphoma cells and breast tumor cells, we compared the expression level of Tiam1 in two human giant-cell lung carcinoma cell strains with high or low metastasis potential, and found that Tiam1 expression level in high-metastatic 95D cells was higher than that in low-metastatic 95C cells. To further confirm the role of Tiam1 in invasion and metastasis, we constructed the antisense Tiam1 expression plasmid (pcDNA3-anti-Tiam1), which was transfected into 95D cells.

A novel gene delivery system targeting urokinase receptor.

Recombinant proteins that combine different functions required for cell targeting and intra-cellular delivery of DNA present an attractive approach for the development of nonviral gene delivery vectors. Here, we described a novel protein termed ATF-lys10 which facilitated cell-specific gene transfer via receptor-mediated endocytosis. ATF-lys10 was composed of the amino-terminal fragment of urokinase and ten lysines at the carboxyl terminus.

Transcriptional regulation of urokinase receptor in high- (95D) and low-metastatic (95C) human lung cancer cells.

To study the transcriptional regulation of urokinase receptor (uPAR) in high- (95D) and low-metastatic (95C) human lung cancer cells, we performed PCR to amplify 2238 bp uPAR promoter from 95C and 95D cells. According to the results of sequencing, five different bases are found in uPAR promoter between 95C and 95D cells. The results of luciferase activity assay showed that these differences have no significant effect on the uPAR promoter activity.

Interaction of plasminogen activator inhibitor-2 and proteasome subunit, beta type 1.

The apoptosis protection by plasminogen activator inhibitor -2(PAI-2) is dependent on a 33 amino acids fragment between helix C and D of PAI-2 which is probably may be due to the interaction of PAI-2 with unknown intracellular proteins. In this study we used the fragment between helix C and D of PAI-2 as bait to screen a HeLa cells cDNA library constructed during apoptosis in a yeast two-hybrid system and retrieved a clone that encodes 241 amino acids of proteasome (prosome, macropain) subunit, beta type 1(PSMbeta1) which plays important roles in NF-kappaB activation. GST-pulldown experiments confirmed the interaction between PAI-2 and PSMB1 in vitro.

[Surgical techniques for the ureterointestinal anastomosis in continent urinary diversion].

Objective: To compare the complications of direct and antirefluxing techniques of ureterointestinal anastomosis in continent urinary diversion.

Methods: Sixty-three patients underwent continent urinary diversion. Twenty-four patients were treated by the direct ureteroenteric anastomosis and the others treated by the antirefluxing technique.

[Application of multihole U-shaped drainage stent in complex posterior urethra surgery].

Objective: To study a method for using a new drainage stent following complex posterior urethral operation.

Methods: Fifty-five patients,15 of whom had complex posterior urethrorectal fistula, 35 had complex posterior stricture or atresia, and 5 had bladder exstrophy, received surgical treatment, after which multihole U-shaped drainage stent was applied.

Results: All the patients were normal in micturition and no complications occurred during the follow-up period lasting for 1 to 10 years.

Identification of interaction between PAI-2 and IRF-3.

HeLa cells transfected with plasminogen activator inhibitor-2 ( PAI-2 ) were protected from TNF- alpha-induced apoptosis. The apoptosis protection by PAI-2 is dependent on a 33 amino acids fragment between helix C and D of PAI-2, which may be due to the interaction of PAI-2 with some intracellular proteins. In this study, the yeast two-hybrid system was used to screen a HeLa cells cDNA library constructed during apoptosis with the fragment between helix C and D of PAI-2 as bait.

[Biological function of fusion protein ATF-PAI2CD].

To express the fusion protein ATF-PAI2CD (urokinase-type plasminogen activator amino terminal fragment-plasminogen activator inhibitor type 2 with the region inter C and D helices deleted ) gene in E.coli and determine the biological characterization of fusion protein ATF-PAI2CD, the cDNA fragment encoding ATF-PAI2CD was cloned into the expression vector pLY-4 and transformed into E.coli JF1125.

[Screening of proteins interacting with tTG in HeLa cells].

Tissue transglutaminase(tTG) belongs to a class of transglutaminase family which is up-regulated in almost all cells apoptosis and is thought to be closely related to cell apoptosis. To investigate the mechanism of tTG in cell apoptosis, yeast two hybrid system was used to screen HeLa cDNA library. One of the 17 positive clones we have obtained encoded the glutamine-rich carboxyl terminus of TIAR, and this interaction between tTG and TIAR, which was finely regulated by Ca(2+), was proved in vitro by GST pull-down.

Interventional therapy for stenosis and aneurysm in renal graft artery: report of 9 cases.

Objective: To study the interventional therapy for renal graft artery stenosis and aneurysm patients with renal transplantation to further improve the survival rate of the graft.

Method: Seven patients with of renal graft artery stenosis received balloon dilatation of the stenotic artery, followed by stent implantation. For renal graft artery aneurysm in another 2 patients, thrombin infusion and stent implantation were respectively performed.

[Misdiagnosis of corticomedullary mixed pathological changes in adrenals: report of 4 cases].

Pathological changes usually occur independently in the adrenal cortex and medulla because of their distinct embryonic origins, and changes involving both the cortex and medulla are rare. We report 4 cases of corticomedullary mixed pathological changes adrenal glands. CT scanning of the adrenal glands showed unilateral abnormalities in all the 4 cases, 3 of which were diagnosed as aldosteronism and the other pheochromocytoma before surgery.

[Diagnosis and treatment of adrenal medullary hyperplasia (with report of 8 cases)].

Objective: To study the diagnosis and treatment of adrenal medullary hyperplasia (AMH).

Methods: An retrospective analysis of the clinical data of 8 patients with AMH admitted in our hospital from May 1998 to May 2002 were conducted with a review of the follow-up study.

Results: CT scanning of the adrenal gland showed unilateral abnormal appearance in all 8 cases.

The Protection Function of the PAI-2 from TNF-alpha is Dependent upon Its Protein Binding Domain.

The cDNA of PAI-2 mutants, PAI-2CD and PAI-2Q were inserted into eukaryotic expression vector, pcDNA3, producing pZLE-PAI2CD and pZLE-PAI2Q respectively. The PAI-2 mutant expression plasmids were transfected into HeLa cells and the transfected cells expressing mutant proteins were selected by G418, Northern blot and ELISA assay. The ELISA assay showed that the expression level of PAI-2 mutant protein was equal to that of wild type PAI-2 in transfected cells.

Biochemical Characterization of PAI-2 and Its Mutants.

To study and compare the biochemical characterization of PAI-2 and its mutants, PCR and site-directed mutagenesis methods were used to generate two PAI-2 mutants, PAI-2CD and PAI-2Q, respectively. The two mutant cDNAs were inserted into prokaryotic expression vector and expressed in a special strain of E.coli, JF1125.

Comparative Studies on the Functions and Characteristics of the Glycosylated, the Nonglycosylated and the Recombinant PAI-1.

Glycosylated and nonglycosylated forms of PAI-1 were separated from HepG 2 cell by immunoaffinity chromatography with anti-PAI-1 monoclonal antibody, and the recombinant PAI-1 were purified from the media of expressed bacteria line of pYZhBI-66. Some functions and characteristics of the three PAI-1 were compared, such as the inhibiting effect on tPA, the activation by denaturation, the stability to thermal or pH changes, and the activation by fibrinogen and heparin. The results emphasized the roles of the oligosaccharide.