Objective To investigate the effect of a newly developed photosensitizer—chlorophyll derivative combined with irradiation of 650 nm laser for PC3, an androgen independent cell line in vitro. Methods PC3 was cultured and designed to 4 groups,including blank control,laser irradiation,medication of photosensitizer and medication of photosensitizer with laser irradiation (treated).The medicated concentration of chlorophyll was 0.1 g/L and irradation fluence of 650 nm semiconductor laser was 6 J/cm2.Intracellular distribution of photosensitizer and cellular morphological alterations were studied through light microscopy, electron microscopy and laser confocal microscopy. Results It showed the shrinkage, round-up and membrane integrity of treated PC3 under light microscopy.Sable deposits were observed in cytoplasm of cells in both photosensitizer and treated groups.Transmission electron microscopy showed the fragmentation of DNA and condensation of chromatin beneath the karyolemma in treated cells.In cytoplasm,the endoplasmic reticulum and mitochondria swell to form vesicles and vacuoles.It showed the strong red fluorescence in the cytoplasm of treat cells compared with the red fluorescence indifferent to the background through laser confocal microscopy. Conclusions Chlorophyll derivative based photodynamic therapy is able to induce apoptosis of PC3 in vitro.Mitochondria is presumed to be the primary target of photodynamic therapy and trigger the apoptotic pathway.

Objective To study the role of Med19 in bladder cancer by analyzing the effects of lentivirus-mediated suppression of Med19 expression on T24 bladder cancer cells in vitro.Methods The lentivirus vectors containing a small hairpin RNA (shRNA) to target Med19 were constructed.After T24 bladder cancer cells were infected,real-time PCR and Western-blotting were used to study the Med19 expressions in the CON group (non-infected cells),the NC group (Lv-NC-infected cells) and the KD group (Lv-shMed19-infected cells).The influence of Med19 on the proliferation of bladder cancer cells were assessed using MTT,BrdU,colony formation assay and tumorigenicity experiment in mice.Cell cycle was analyzed with flow cytometry assay.Results Med19 relative mRNA level (0.35 ± 0.03) and Med19 protein expressing in the KD group were significantly inhibited (P ＜ 0.05).The KD group displayed an increased proportion of cells (77.50 ± 0.29)％ in the G0/G1 phase compared with the CON group (69.81 ± 0.81)％and NC group (67.53 ± 0.67) ％ (P ＜ 0.05).Compared with the CON group and the NC group,the KD group displayed a significant cell proliferation defect by MTT and BrdU assay and the number of colonies (91.33 ± 6.11) was significant decreased (P ＜ 0.05).On the day 24,the tumor volume (596.64 ± 485.36) mm3 and weight (0.57 ± 0.44) g of the KD group mice were decreased after inoculation into nude mice (P ＜ 0.05).Specific lentivirus-mediated knockdown of Med19 significantly impacted the cell cycle and proliferation of bladder cancer cells.Infected T24 cells nearly lost their tumorigenicity when being inoculated into nude mice.Conclusion Our results provide new evidence of an important role for Med19 in the development of bladder cancer,suggesting that lentiviruses delivering shRNA against Med19 may be a promising tool for bladder cancer therapy.

Objective To study the clinical significance of the mRNA expression level of a novel gene which encodes a kind of transmembrane prostate protein induced by androgen-PMEPA1, as it may predict the progress of prostate cancer from hormone-dependent to hormone-independent. Methods We used Real-time PCR to detect the mRNA expression of PMEPA1 and GSTP1 in prostate cancer cell lines (LNCaP, PC-3), epithelia cells of benign prostatic hyperplasia and tissues from 33 patients with prostate cancers and 16 cases of prostatic hyperplasia. Results We found the mRNA expression of GSTP1 and PMEPA1 were both down-regulated in prostate cancer cell lines. The mRNA expression of GSTP1 was up-regulated in 6.1% of cases, down-regulated in 81.8%, and showed no difference in 12.1%. While PMEPA1 was highly expressed in 27.3% of cases, lowly expressed in 27.3%, and not differently expressed in 45.4%. Statistical analysis showed that the mRNA expression of GSTP1 was relevant to ages, but had no relationship with PSA, TNM stage, osseous metastasis or tumor differentiation, while the mRNA expression of PMEPA1 was relevant to osseous metastasis and tumor differentiation, but had no relationship with age, PSA or TNM stage. Conclusions PMEPA1 is possibly a useful biomarker, as it can identify patients with unfavourable prognosis, however, this hypothesis needs to be further studied with large samples.

AIM: Previous studies have suggested that big stick design (BSD) method can only be used in clinical trials of two treatments with equal proportion, which has good statistical performance and has become the recommended choice of randomized methods. This study expands BSD method, so that it can be applied to three groups, and provides more randomized methods for clinical trials. METHODS: On the basis of BSD method used in two treatments with equal proportion, the derivation conditional allocation probability of BSD method used in three treatments with equal proportion was carried out. BSD method was compared with simple randomization (SR) method, permuted block design (PBD) method and block urn design (BUD) method by Monte-Carlo simulation in balance and randomness. RESULTS: In terms of balance, PBD method was the best, followed by BUD method, BSD method, and SR method was the worst. In terms of randomness, SR method was the best, followed by BSD method, BUD method and PBD method. The comprehensive performance showed that BSD method was better than BUD method, PBD method and SR method. CONCLUSION: The expanded BSD method used in three treatments with equal proportion has good comprehensive performance, and it can be the recommended randomization method for clinical trials of three treatments with equal proportion.