Zeng Fuqing

Expression and significance of Oct4 in bladder cancer

SummaryIn order to detect the expression of Oct4 in bladder cancer tissue and cell line BIU-87, immunohistochemistry was used in 49 bladder cancer biopsy samples and immunofluorescence and reverse transcription-PCR were performed on bladder cancer cell line BIU-87. Forty of 49 bladder cancer samples showed the expression of Oct4 in about 0.6% cancer cells. The positive rate and density of Oct4 expression had no obvious relationship with the grade, recurrence or metastasis of bladder cancer (P>0.05). A few Oct4 positive cells were found in bladder cancer cell line BIU-87, which was also confirmed by RT-PCR. This study indicated the existence of few Oct4 positive cells in bladder cancer, which may be the bladder cancer stem cells. This study may provide the foundation for isolation and identification of bladder cancer stem cells.In order to detect the expression of Oct4 in bladder cancer tissue and cell line BIU-87, immunohistochemistry was used in 49 bladder cancer biopsy samples and immunofluorescence and reverse transcription-PCR were performed on bladder cancer cell line BIU-87. Forty of 49 bladder cancer samples showed the expression of Oct4 in about 0.6% cancer cells. The positive rate and density of Oct4 expression had no obvious relationship with the grade, recurrence or metastasis of bladder cancer (P>0.05). A few Oct4 positive cells were found in bladder cancer cell line BIU-87, which was also confirmed by RT-PCR. This study indicated the existence of few Oct4 positive cells in bladder cancer, which may be the bladder cancer stem cells. This study may provide the foundation for isolation and identification of bladder cancer stem cells.

Preparation of curcumin prodrugs and theirin vitro anti-tumor activities

SummaryThe curcumin prodrugs, which could be selectively activated in tumor cells, were prepared to establish a basis for the targeted chemotherapy for cancer. On the basis of the molecular structure of curcumin, the N-maleoyl-L-valine-curcumin (NVC), N-maleoyl- glycine-curcumin (NGC) were chemically synthesized and identified by IR and NMR spectroscopy. After treatment with these two prodrugs for 6–24 h, the rates of growth inhibition on human bladder cancer EJ cells and renal tubular epithelial (HKC) cells were detected by MTT colorimetry. Our results showed that after the treatment with 20 μmol/L–40 μmol/L NVC and NGC for 6–24 h, the growth inhibitory effects on EJ cells were 6.71%–65.13% (P<0.05), 10.96%–73.01% (P<0.05), respectively, in both dose- and time-dependent manners. When compared with the curcumin of same concentrations, the growth inhibitory effects of these two prodrugs on HKC cells were significantly decreased (P<0.01). It is concluded that activation of curcumin prodrugs via hydrolysis functions of cellular esterase could inhibit the growth activities of tumor cells, and reduce the side effects on normal diploid cells. This provided a novel strategy for further exploration of tumor-targeted chemotherapeutic drugs.

Expression of beta-human chorionic gonadotropin genes in renal cell cancer and benign renal disease tissues.

SummaryTo study the expression of beta-human chorionic gonadotropin (βhCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of βhCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues. It was found that 52% RCC samples revealed positive for βhCG mRNA expression. Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference. The positive rate of βhCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys. β7 was most frequently transcribed subtype gene independent on the histology. These findings suggested βhCG gene transcription is not only involved in RCC but also in benign renal diseases.

Smac/DIABLO promotes mitomycin C-induced apoptosis of bladder cancer T24 cells

SummaryThe enhancing effects of Smac gene on the mitomycin C-induced apoptosis of the bladder cancer cell line T24 were investigated. The Smac gene was transfected into bladder cancer cell line T24 under the induction of liposome. The intrinsic Smac level was detected by using immunohistochemistry and RT-PCR. The in vitro cellular growth activities were assayed by MTT colorimetry. Apoptosis was assayed by the flow cytometry. The results showed that as compared with the control cells, the apoptosis rate of T24 cells induced by mitomycin C was enhanced by transfected Smac gene. Flow cytometry revealed that, the apoptosis rate was 18.84% and 33.52%, and 10.72% and 26.24% respectively in blank and transfected cells treated with 0.05 or 0.005 mg/mL mitomycin C (P<0.05). It was concluded that Smac could enhance the apoptosis of T24 by mitomycin C, which could provide a useful experimental evidence for bladder cancer therapy.

Expression of X-linked inhibitor of apoptosis protein and its effect on chemotherapeutic sensitivity of bladder carcinoma

The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P<0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P<0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P<0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemotherapeutic sensitivity of T24 cells.

Preparation of Arsenic Trioxide Albumin Microspheres and its Release Characteristics in Vitro

SummaryArsenic trioxide albumin microspheres (As2O3-BSA-NS) were prepared by using methods of chemical cross-linking. The desirability function (DF), calculated according to the size (<1μm) distribution, drug loading and drug trapping efficiency, was introduced as a total index for the microspheres formulation. Four factors, inculding W/O ratio, decentralization speed, BSA concentration and stirring stabilization time, were selected and arranged in an orthogonal experimental table. The release characteristic was studied by the drug release experimentin vitro. The four factors affected DF differently. Decentralization speed behaved as the maximum (P<0.01), followed by BSA concentration (P<0.05) and the W/O ratio dose (P<0.05). Stirring stabilization time did not influence DF (P>0.05). The release experimentin vitro showed that As2O3 in As2O3-BSA-NS was released more slower than pure As2O3. It was concluded that regular As2O3-BSA-NS may be prepared by the methods of chemical cross-linking, which was optimized by orthogonal experimental analysis of different factors, and the microspheres can release As2O3 slowly.

The relationship of expression of bcl-2, p53, and proliferating cell nuclear antigen (PCNA) to cell proliferation and apoptosis in renal cell carcinoma.

SummaryTo investigate the relationship of bcl-2, p53, proliferating cell nuclear antigen (PCNA) to cell proliferation, apoptosis and pathological parameters, the patterns of cell growth and turnover in renal cell carcinoma (RCC), formalin-fixed and paraffin-embedded tissue blocks from 34 patients with RCC were examined. Cell proliferation activity was detected by PCNA immunostaining and the proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy-nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL-positive cells in the tumor cells. Expressions of bcl-2 and p53 were assessed immunohistochemically. Our results showed that the PI ranged from 6.0% to 24.0% (median 12. 3%) and the AI from 2.0% to 8.0% (median 5.4%) in RCC. The expression of the bcl-2 protein was demonstrated in 15 cases (44.1%); the expression of the p53 protein, however, was seen in only 3 case. bcl-2 positivity was not associated with PI or AI or any pathological parameters. There were close associations between PI and tumor grade and stage, and a significant relationship between AI and the tumor grade of RCC. Our study suggests that bcl-2 positivity was not associated with PI or AI or any pathological parameters. There are close associations between PI and AI and tumor grade and stage of RCC. Active cell proliferation may be accompanied by frequent apoptosis in RCC.

Ureteroscopic holmium: YAG laser lithotripsy for managing ureteral calculi (A report of 168 cases)

The effectiveness and safety of ureteroscopic holmium: YAG laser lithotripsy for managing ureteral calculi was evaluated. Ureteroscopic holmium. YAG laser lithotripsy was performed in 168 ureteral calculi (upper 27 cases, middle 33 cases and lower 108 cases). The results showed that the stone-free rate was 92.6% in the upper ureteral calculi, 93.9% in the middle ureteral calculi and 94.4% in the lower ureteral calculi, respectively. The complication rate was 4.8% (8 cases). It was suggested that ureteroscopic holmium: YAG laser lithotripsy is a highly effective and safe treatment modality for managing ureteral calculi.SummaryThe effectiveness and safety of ureteroscopic holmium: YAG laser lithotripsy for managing ureteral calculi was evaluated. Ureteroscopic holmium: YAG laser lithotripsy was performed in 168 ureteral calculi (upper 27 cases, middle 33 cases and lower 108 cases). The results showed that the stone-free rate was 92.6% in the upper ureteral calculi, 93.9% in the middle ureteral calculi and 94.4% in the lower ureteral calculi, respectively. The complication rate was 4.8% (8 cases). It was suggested that ureteroscopic holmium: YAG laser lithotripsy is a highly effective and safe treatment modality for managing ureteral calculi.

Usefulness of real-time three-dimensional ultrasonography in percutaneous nephrostomy: an animal study

To evaluate the effect of real‐time three‐dimensional (3D) ultrasonography (US) in guiding percutaneous nephrostomy (PCN).

Cloning of human Uroplakin II gene from Chinese transitional cell carcinoma of bladder and construction of its eukaryotic expression vector

SummaryTo clone Uroplakin II gene from Chinese transitional cell carcinoma (TCC) of bladder and construct its eukaryotic expression vector, the molecular cloning method was used to extract total RNA from a GIII/T3N0M0 tissue sample of the bladder TCC patients. The primers were designed by Primer 5.0, software. Full length cDNA of Uroplakin II gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), assayed by nucleic acid sequencing and then inserted betweenXba I andHindIII restrictive sites of eukaryotic expression vector pcDNA3.0. The recombinant was assayed by restricted enzyme digestion. Under the induction of Lipofectamine 2000, the recombinant was transfected into Uroplakin II negative bladder cancer cell line EJ. Cellular expression levels of Uroplakin II were detected by RT-PCR. The nucleic acid sequencing results indicated that Chinese Uroplakin II cDNA (555 bp) was successfully cloned. The BLAST analysis demonstrated that the cloned sequence is 100% homologous with sequences reported overseas. The GenBank accession number AY455312 was also registered. The results of restricted enzyme digestion indicated that eukaryotic vector pcDNA-UP II for Uroplakin II, was successfully constructed. After being transferred with pcDNA-UP II for 72 h, cellular Uroplakin II mRNA levels were significantly improved (P<0.01). It is concluded that human Uroplakin II gene was successfully cloned from Chinese TCC tissues, which provided a basis for further exploration of the roles of Uroplakin II gene in TCC biological behaviors and potential strategies for targeted biological therapy of TCC.