Ke-qin Zhang

University of South China, Heng-nan, Hunan, China

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Publications (9)14.92 Total impact

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    ABSTRACT: TNF-α and IFN-γ are the major pro-inflammatory cytokines in the β-cell destruction. However, the underlying mechanism remains unclear. The present study used a murine insulinoma cell line MIN6 for further investigation of the effect of Caspase-3 on the cytokines-induced pancreatic β-cell apoptosis and analyzed the mechanisms involved in the activation of Caspase-3. It was showed that the combination of IFN-γ and TNF-α significantly reduced the viability of MIN6 cells and the observed cells growth inhibition was due to cell apoptosis as judged by the morphological changes under a confocal laser scanning microscopy and FACS assay of Annexin-V/7-AAD double staining. Accompanying with NF-κB activation and Bcl-2 downregulation, both the cleaved Caspase-3 and PARP, a known substrate of Caspase-3 in vivo, were observed at 24 and 12 h, respectively, after cells exposure to IFN-γ and TNF-α treatment. Pretreatment of Caspase-3 inhibitors remarkably attenuated IFN-γ- and TNF-α-induced cells apoptosis. Inhibition of NF-κB activation led to the increase in Bcl-2 expression, a significant attenuation in Caspase-3 activity, and an obvious amelioration in cells viability in IFN-γ- and TNF-α-treated MIN6 cells. Taken together, our results indicate that Caspase-3 is critical for the induction of MIN6 cells apoptosis and it's activation is further confirmed to be related to the NF-κB-mediated Bcl-2 downregulation, which may be the underlying mechanism of IFN-γ- and TNF-α-mediated MIN6 cells apoptosis.
    Cell biochemistry and biophysics 05/2013; · 3.34 Impact Factor
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    ABSTRACT: To develop a completely novel DNA peptide-combined vaccine and determine whether it can efficiently improve tumor-specific cytotoxic T lymphocyte (CTL) responses and inhibit tumor progression in experimental prostate cancer models. The DNA/peptide combined vaccine was prepared by the self-assembly of a cationic peptide ([K]18P9) containing 18 lysines and a CTL epitope peptide, prostate stem cell antigen (PSCA (14-22)) (HLA-A2 restricted) with a recombinant plasmid encoding human full-length PSCA gene (pcDNA3.1(+)-PSCA) through electrostatic interactions. The formation of a DNA/peptide complex was examined by DNA retardation assay, DNase I protection assay, and transmission electron microscopy. The efficacy of vaccination using this complex was demonstrated in terms of the PSCA-specific CTL activity and antitumor immunity to PSCA(+) tumors in a murine model. This form of DNA/peptide complex could efficiently transfer the plasmid encoding full-length PSCA gene into mammalian cells and induced potent CTLs cytotoxicity against a human prostate carcinoma cell line established from the left supraclavicular lymph node metastasis from a 50-year-old man with prostate carcinoma in 1977. Expressing PSCA compared with pcDNA3.1(+)-PSCA, [K]18P9 peptide, or pcDNA3.1(+). Moreover, the vaccination of mice with this complex induced a potent antitumor immunity to prostate carcinomas in a xenograft tumor model in nude mice. This study suggests that a specific antitumor immune response can be induced by this DNA/peptide combined vaccine, which represents a new strategy for use in the immunotherapy of prostate cancer.
    Urology 04/2012; 79(6):1410.e7-13. · 2.42 Impact Factor
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    ABSTRACT: To report our preliminary techniques and experience with transumbilical laparoendoscopic single-site renal pedicle lymphatic disconnection (LESS-RPLD) in seven patients with refractory chyluria. Between June 2009 and September 2010, seven patients with refractory chyluria underwent LESS-RPLD. In the patients, a 2- to 3-cm single inverted U-shaped supraumbilical incision was made, and a homemade single multichannel port using a surgical glove and three conventional trocars was placed into the abdominal cavity. Flexible electric coagulation hook and pliers were used for renal pedicle dissection. A straight ultrasound knife was used for lymphatic disconnection. All the operations were successfully completed without conversion to open surgery, although an additional 3-mm trocar was used to push the liver in one patient. The mean operative time was 125 (96-165) minutes. The mean blood loss was estimated to be 112 (50-250) mL. Chyluria disappeared in all patients after surgery and did not recur during the follow-up period (3-15, mean 8.3 mos). LESS-RPLD is safe and feasible, with favorable short-term outcomes and aesthetic effect.
    Journal of endourology / Endourological Society 08/2011; 25(8):1337-41. · 1.75 Impact Factor
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    ABSTRACT: Although the critical role of complement component C3d as a molecular adjuvant in preventing virus infection is well established, its role in cancer therapies is unclear. In this study, we have engineered a DNA vaccine that expresses extracellular region of murine VEGFR-2 (FLK1(265-2493)) and 3 copies of C3d (C3d3), a component of complement as a molecular adjuvant, designed to increase antitumor immunity. VEGFR-2 has a more restricted expression on endothelial cells and is upregulated once these cells proliferate during angiogenesis in the tumor vasculature. Immunization of mice with vector encoding FLK1(265-2493) alone generated only background levels of anti-VEGFR-2 antibodies and slight inhibitory effect on tumor growth. However, the addition of C3d3 to the vaccine construct significantly augmented the anti-VEGFR-2 humoral immune response and inhibited the tumor growth. The antitumor activity induced by vaccination with vector encoding FLK1(265-2493)-C3d3 fusion protein was also demonstrated via growth inhibition of established tumors following passive transfer of immune serum from vaccinated mice. Our results suggest that vaccination with vector encoding FLK1(265-2493) with C3d3 as a molecular adjuvant induces adaptive humoral activity, which is directed against the murine VEGFR-2 and can significantly inhibit tumor growth, and that administration of C3d as a molecular adjuvant to increase antibodies levels to VEGFR-2 may provide an alternative treatment modality for cancer therapies.
    Vaccine 10/2010; 28(44):7221-7. · 3.77 Impact Factor
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    ABSTRACT: To evaluate the one-hole method for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy. We used the one-hole method for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy in the treatment of 15 cases of varicocele, 13 in the left and 2 in the right side, and appraised the treatment results by follow-up 1 month after the surgery. All the operations succeeded and no complications developed. The average operation time was 28 minutes and the mean hospital stay was 4 days. Symptoms were significantly relieved in all the patients and the scars were inconspicuous at follow-up. The one-hole method is a novel option for high ligation of the internal spermatic vein by embryonic natural orifice transumbilical laparoscopy in the treatment of varicocele. It is recommendable for its advantages of simple procedure, less pain, few complications, quick recovery, mini-invasiveness and cosmetic acceptability.
    Zhonghua nan ke xue = National journal of andrology 05/2010; 16(5):450-2.
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    ABSTRACT: Although the critical role of complement component C3d as a molecular adjuvant in preventing virus infection is well established, its role in cancer prophylaxis and treatment is unclear. In this study, we constructed a recombinant plasmid encoding Flk-1 and C3d3 fusion proteins and investigated its transient expression in vitro in transfected eukaryotic cells and its antibody response in immunized mice. Subsequently, we investigated the vaccine's ability to elicit an immune response leading to suppression of angiogenesis and tumor growth in mice bearing bladder transitional cell carcinoma. Using Western blotting, immunocytochemistry, and flow cytometry, we detected the expression of Flk-1 and C3d3 fusion proteins in COS-7 cells transfected with these recombinant plasmids. Further binding experiment using CR2 (C3d receptor) positive Raji cells that were incubated with transfected COS-7 supernatant indicated that C3d was successfully fused to Flk-1. Although both vaccines elicited peak antibody levels at 5 weeks, Flk-1-specific antibody titer in pSG.SS.Flk-1(ECD).C3d3.YL-immunized mice was significantly higher when compared to pSG.SS.Flk-1(ECD).YL-immunized mice. The results of experiments with bladder tumor-bearing mice showed that the vaccine inhibited tumor growth significantly. These results suggest that C3d plays a critical role in tumor immunotherapy by promoting antibody response in Flk-1-based DNA vaccines. This approach may provide a new strategy for the rational design of anti-angiogenic therapies for the treatment of solid tumors and provide a basis for the further exploitation and application of the anti-angiogenesis DNA vaccines.
    Cancer Immunology and Immunotherapy 07/2009; 59(1):93-101. · 3.64 Impact Factor
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    ABSTRACT: To review retrospectively the urological complications in 1 223 kidney transplants. A total of 1 223 kidney transplants were divided into ureteroneocystostomy group (n = 948) and ureteroureterostomy group (n = 275) according to the methods of urinary tract reconstruction. The incidence and management of urological complications such as urinary fistula, obstruction of ureter, vesicoureteral reflux (VUR) and urinary tract infection (UTI) were summarized respectively. Overall, urological complications were encountered in 217 (17.7%) cases, including 43 cases of urinary fistula (3.5%), 35 obstruction of ureter (2.9%), 14 VUR (1.1%) and 125 UTI (10.2%). Urinary fistula was 39 (4.1%) cases and 4 cases (1.5%) (P < 0.05), obstruction of ureter 22 (2.3%) & 13 (4.7%) (P < 0.05), VUR 14 (1.5%) & 0 (0%) (P < 0.05) and UTI 109 (11.5%) & 16 (5.8%) (P < 0.01) in the ureteroneocystostomy group and ureteroureterostomy group respectively. Seventy patients underwent surgical treatment. The 3-year survival rate of graft with urological complications and without urological complications were 82.3% and 84.7% respectively. Ureteroureterostomy can decrease the incidence of urological complications after kidney transplantation. Most of urological complications require surgical interventions. The long-term graft survival is not affected by a correctly treated urological complication.
    Zhonghua yi xue za zhi 05/2009; 89(18):1269-71.
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    ABSTRACT: To evaluate the effect of testis homotransplantation in the treatment of androgen deficiency and infertility. We retrospectively analyzed 12 cases of testis homotransplantation. Surgical success was achieved in 11 cases, all with a significantly increased level of serum testosterone, and markedly improved secondary sex characteristics and sexual function. Testis homotransplantation is highly effective for the treatment of androgen deficiency in males, but has little effect on spermatogenesis.
    Zhonghua nan ke xue = National journal of andrology 04/2008; 14(3):248-50.
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    ABSTRACT: To explore the specific surface markers for the isolation and purification of human spermatogonial stem cells (SSC). Specific markers of human SSC were screened and identified in fetal and adult testes by immunohistochemical assay, using HSC markers c-kit, Thy-1 and human ES integrins. In human adult testes, the alpha6 integrin extensively and significantly expressed on the surface of most of the germ cells in the seminiferous tubule, and beta1 integrin mainly expressed on the surface of the germ cells residing on or near the basal membrane in the seminiferous tubule. Thy-1 scattering expressed on the surface of some cells of the basal membrane, and on some Leydig cells as well. The three antigen markers expressed on the SSC of human adult testes specifically to some extent. SSEA-1 specifically expressed on the surface of the gonocytes in the fetal testes. The alpha6 and beta1 integrins and Thy-1 may be used for the SSC isolation as positive markers. SSEA-1 can be used as an identification marker for the fetus SSC.
    Zhonghua nan ke xue = National journal of andrology 08/2005; 11(7):486-9.