It has been widely verified by various sorting methods that cancer stem cells (CSCs) exist in different types of tumor cells or tissues. However, due to lack of specific stem cell surface markers, CSCs are very difficult to be separated from some cancer cells, which becomes the key barrier of functional studies of CSCs. The sorting method by side population cells (SP) lays a solid foundation for in-depth and comprehensive study of CSCs. To identify the existence of SP in prostate cancer cell lines, we applied flow cytometry sorting by SP to cultures of prostate cancer cell lines (TSU, LnCap, and PC-3), and the cancer stem-like characteristics of SP were verified through experiments in vitro and in vivo. The proportion of SP in TSU cells was calculated to be 1.60%±0.40% [Formula: see text], and that in PC-3 and LnCap cells was calculated to be 0.80%±0.05% and 0.60%±0.20%, respectively. The colony formation assay demonstrated that the colony formation rate of SP to non-SP sorted from TSU via flow cytometry was 0.495±0.038 to 0.177±0.029 in 500 cells, 0.505±0.026 to 0.169±0.024 in 250 cells, and 0.088±0.016 to 0.043±0.012 in 125 cells respectively. In the in vivo experiments, tumors were observed in all the mice on the 10th day after injecting 50 000 cells subcutaneously in SP group, whereas when 5×10(6) cells were injected in non-SP group, tumors were developed in only 4 out of 8 mice until the 3rd week before the end of the experiment. Our results revealed that prostate cancer cells contain a small subset of cells, called SP, possessing much greater capacity of colony formation and tumorigenic potential than non-SP. These suggest that SP in prostate cancer cells may play a key role in the self-renewal and proliferation, and have the characteristics of cancer stem-like cells. Dissecting these features will provide a new understanding of the function of prostate CSCs in tumorigenicity and transformation.

Objective To investigate the safety and efficacy of photoselective green laser vaporization of bladder tumor (PVBT). Methods A total of 522 patients with bladder tumor were enrolled in present study from January 2010 to May 2015, including 405 cases of non muscle-invasive bladder cancer (NMIBC) and 117 cases of muscle-invasive bladder cancer (MIBC). All of patients were treated with PVBT and intravesical instillation of epirubicin. Patients with MIBC received intravenous chemotherapy (kisi-hama and cisplatin). Results The hospitalization time was (7.32±1.28) days, the operation time was (27.08±5.36) min, and the indwelling urinary catheter was (2.42±0.34) days for patients in NMIBC group. During the follow-up period (12-60 months), 38 cases (9.4％) relapsed, of which 3 cases underwent radical cystectomy, and other 35 cases underwent PVBT again. All 405 patients were alive at the end of follow-ups. The hospitalization time was(26.18 ± 1.92) days, the operation time was (38.32 ± 6.54) min, and the time of indwelling urinary catheter was (2.72 ± 0.85) days for patients of MIBC group. During the follow-up period (12-60 months), 19 cases (16.2％) relapsed. Among them, 4 patients underwent radical cystectomy, and other 15 cases underwent PVBT. Six patients died from distant organ metastasis (including 2 cases of pulmonary metastasis and 4 cases of bone metastasis), and other 111 patients survived. Conclusion PVBT is safe and effective in the clinical application, especially for NMIBC and MIBC patients who are unable or unwilling to undergo radical cystectomy.

It has been widely verified by various sorting methods that cancer stem cells (CSCs) exist in different types of tumor cells or tissues. However, due to lack of specific stem cell surface markers, CSCs are very difficult to be separated from some cancer cells, which becomes the key barrier of functional studies of CSCs. The sorting method by side population cells (SP) lays a solid foundation for in-depth and comprehensive study of CSCs. To identify the existence of SP in prostate cancer cell lines, we applied flow cytometry sorting by SP to cultures of prostate cancer cell lines (TSU, LnCap, and PC-3), and the cancer stem-like characteristics of SP were verified through experiments in vitro and in vivo. The proportion of SP in TSU cells was calculated to be 1.60%±0.40% [Formula: see text], and that in PC-3 and LnCap cells was calculated to be 0.80%±0.05% and 0.60%±0.20%, respectively. The colony formation assay demonstrated that the colony formation rate of SP to non-SP sorted from TSU via flow cytometry was 0.495±0.038 to 0.177±0.029 in 500 cells, 0.505±0.026 to 0.169±0.024 in 250 cells, and 0.088±0.016 to 0.043±0.012 in 125 cells respectively. In the in vivo experiments, tumors were observed in all the mice on the 10th day after injecting 50 000 cells subcutaneously in SP group, whereas when 5×10(6) cells were injected in non-SP group, tumors were developed in only 4 out of 8 mice until the 3rd week before the end of the experiment. Our results revealed that prostate cancer cells contain a small subset of cells, called SP, possessing much greater capacity of colony formation and tumorigenic potential than non-SP. These suggest that SP in prostate cancer cells may play a key role in the self-renewal and proliferation, and have the characteristics of cancer stem-like cells. Dissecting these features will provide a new understanding of the function of prostate CSCs in tumorigenicity and transformation.
Cell Line, Tumor ; Humans ; Male ; Neoplastic Stem Cells ; pathology ; Prostatic Neoplasms ; pathology ; Side-Population Cells ; pathology

Objective To investigate the effect of adjuvant therapy of Shenfu Injection on NOD-like receptor protein 3(NLRP3)/cysteine-aspartic acid-specific protease 1(Caspase-1)mediated py-roptosis signaling pathway and inflammatory factor levels in rats with acute myocardial infarction(AMI).Methods A total of 40 rats were randomly divided into sham operation group,model group,betaloc group(0.9 mg/kg),and combination group(0.9 mg/kg betaloc combined with 6 mL/kg Shenfu Injection),with 10 rats in each group.The rats were treated by gavage continuously for 3 weeks.The levels of serum troponin Ⅰ(cTnⅠ),creatine kinase-MB(CK-MB),interleukin(IL)-6,IL-1β,and tumor necrosis factor-α(TNF-α)in rats were detected before modeling,after modeling,and at 3 weeks of treatment.Echocardiographic parameters such as left ventricular ejection fraction(LVEF),left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),and myocardial infarction area were compared among groups after modeling and at 3 weeks of treatment.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot were used to detect the mRNA and protein expression levels of NLRP3 and Caspase-1 in the myocardium.Results Compared with the sham operation group,the levels of cTnⅠ,CK-MB,IL-6,IL-1 β,TNF-α,myocardial infarction area,and the expressions of NLRP3 mRNA and Caspase-1 mRNA in the model group were significantly increased after modeling and at 3 weeks of treatment(P＜0.05);compared with the model group,the levels of cTnⅠ,CK-MB,IL-6,IL-1β,TNF-α,myocardial infarction area,and the expressions of NLRP3 mRNA and Caspase-1 mRNA in the betaloc group and the combination group were significantly decreased at 3 weeks of treatment,and the above indicators in the combination group were significantly lower than those in the betaloc group(P＜0.05).Conclusion Adjuvant therapy of Shenfu Injection for AMI can fur-ther alleviate myocardial cell injury,shorten infarction size,and inhibit inflammatory response and pyroptosis activity.

Objective To investigate the effect of adjuvant therapy of Shenfu Injection on NOD-like receptor protein 3(NLRP3)/cysteine-aspartic acid-specific protease 1(Caspase-1)mediated py-roptosis signaling pathway and inflammatory factor levels in rats with acute myocardial infarction(AMI).Methods A total of 40 rats were randomly divided into sham operation group,model group,betaloc group(0.9 mg/kg),and combination group(0.9 mg/kg betaloc combined with 6 mL/kg Shenfu Injection),with 10 rats in each group.The rats were treated by gavage continuously for 3 weeks.The levels of serum troponin Ⅰ(cTnⅠ),creatine kinase-MB(CK-MB),interleukin(IL)-6,IL-1β,and tumor necrosis factor-α(TNF-α)in rats were detected before modeling,after modeling,and at 3 weeks of treatment.Echocardiographic parameters such as left ventricular ejection fraction(LVEF),left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),and myocardial infarction area were compared among groups after modeling and at 3 weeks of treatment.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot were used to detect the mRNA and protein expression levels of NLRP3 and Caspase-1 in the myocardium.Results Compared with the sham operation group,the levels of cTnⅠ,CK-MB,IL-6,IL-1 β,TNF-α,myocardial infarction area,and the expressions of NLRP3 mRNA and Caspase-1 mRNA in the model group were significantly increased after modeling and at 3 weeks of treatment(P＜0.05);compared with the model group,the levels of cTnⅠ,CK-MB,IL-6,IL-1β,TNF-α,myocardial infarction area,and the expressions of NLRP3 mRNA and Caspase-1 mRNA in the betaloc group and the combination group were significantly decreased at 3 weeks of treatment,and the above indicators in the combination group were significantly lower than those in the betaloc group(P＜0.05).Conclusion Adjuvant therapy of Shenfu Injection for AMI can fur-ther alleviate myocardial cell injury,shorten infarction size,and inhibit inflammatory response and pyroptosis activity.

Ureterosciatic hernia is a rare type of pelvic floor herniation with no typical symptoms. The resulting ureteral obstruction may lead to hydronephrosis and complications such as urinary tract infection and urosepsis.Fewer than 40 ureterosciatic hernia cases have been reported all over the world. Active surveillance, manual reduction, ureteral stenting and surgical repair are common treatment options. Among them, the safety and effectiveness of laparoscopic herniorrhaphy have been recognized. This article review the advances in the diagnosis and treatment of ureterosciatic hernia.

With the completion of large-scale genome sequencing of human beings and other organisms, understanding the expression of control elements on the genome has become an important research task in the post-genome era. The enhancer trapping technology is an effective method for identifying enhancer elements in the genome and understanding its mechanism for gene expression regulation. In this study, we selected the stable enhancer trapping line TK4 (head and trunk specific GFP expression), which is generated with the mediation of Tol2 transposon system, and analyzed the trapped enhancers with the techniques of Splinkerette PCR (sp-PCR), in situ hybridization and comparative genomics. We crossed F1 individuals of TK4 line with wild-type zebrafish, collected fertilized eggs, and then detected the expression pattern of green fluorescent protein reporter gene by fluorescence microscopy at six different developmental stages, 6 hpf (hour post fertilization), 24 hpf, 48 hpf, 3 dpf (day post fertilization), 4 dpf and 5 dpf . The zebrafish genome flank sequence near the insertion site of Tol2 transposon was cloned by sp-PCR, and the results revealed that the insertion located at the position 27749253 of chromosome 23, and the transgene inserted reversely inside the intron 1 of rps26 gene. Within the 100 kb region of the insertion site, totally, seven genes including arf3a, wnt10b, wnt1, rps26, IKZF4, dnajc22 and lmbr1l were identified. Comparative genomic analysis by VISTA program revealed that there were two potential enhancer elements in the downstream of rps26 gene, which were conserved non-coding sequence (CNS) 1 and CNS2. The results of in situ hybridization showed that two transcripts of rps26 gene were maternal expression, the expression of rps26-201 in zygote was earlier than that of rps26-001, and the GFP signal of TK4 line zebrafish was not detectable before 6hpf, the expression patterns of rps26 and GFP at the late stages display similarity, and also represent differences, which suggested that the expression of rps26 and GFP may be controlled by the same enhancer, and also by the different enhancer, and two potential enhancers (CNS1 and CNS2) may play a differential regulation roles on the spatial and temporal expression of nearby genes (including rps26). In this study, we successfully obtained two potential enhancers near rps26 gene for the first time, which laid a foundation for further study of the regulation mechanism between these two enhancers and nearby genes in the genome, and the combination technique used in this study also provides a reference for enhancer analysis.