Hyperbaric oxygen therapy (HBOT) is defined as a treatment in which a patient intermittently breathes 100% oxygen while the treatment chamber is pressurized to a pressure greater than sea level (1.0 atmosphere absolute, ATA). In China, for nearly 50 years, HBOT has been used as a primary or adjuvant therapy to treat a variety of diseases. This article mainly reviewed the indications and contraindications of HBOT, as well as the status of clinical and experimental HBOT research in China. At the same time, there is a brief introduction of hyperbaric oxygen preconditioning (HBO-PC) in China.

Transitional cell carcinoma of bladder (TCCB) is a common malignancy worldwide, and outcomes for patients with advanced bladder cancer remain poor. To study the pathogenesis of TCCB, we investigated roles of Phospholipase C (PLC)ɛ, an effector of Ras and Rap small GTPases. RNA interference was used to knockdown PLCɛ expression in human bladder cancer cell lines (BIU-87 and T24). The expression levels of PLCɛ mRNA and protein were detected by reverse transcriptase-polymerase chain reaction and Western blot, respectively. Flow cytometry (FCM) was used to detect distribution of cell cycle. Cellular apoptosis was reflected by transmission electron microscopy and the expression of bcl-2 and bax. We found that PLCɛ could be efficiently knocked down by shRNA. FCM assay showed that the pGenesil-PLCɛ-transfected cells were arrested at the G0/G1 phase. Silence of PLCɛ might induce apoptosis via modulation of bcl-2 and bax. In conclusion, our results suggest that PLCɛ plays an important role in the pathogenesis of human bladder cancer cells. PLCɛ may be used as a potential target of gene therapy for bladder cancer in future.

Objective To investigate the effect of transfection of hepatocyte cell adhesion molecule(HEPACAM) on apoptosis of renal cell carcinoma cells.Methods The vector containing HEPACAM gene was transfeceted into 786-0 cells,the expression of HEPACAM gene in 786-0 cells was confirmed by RT-PCR;The morphological change of nucleus was shown by Hoechst staining;The proliferation and apoptosis of 786-0 cells were analyzed by FCM;The expression of BAX,BCL-2,Cyclin D1 was measured by Western blot.Results Up-regulated expression of HEPACAM gene in 786-0 cells can arrest cell cycle at G0/G1 phase and induce cell apoptosis,the expression of BAX increased and the expression of BCL-2 and Cyclin D1 decreased.Conclusion Ectogenic transfected HEPACAM gene may induce cell apoptosis and arrest cell cycle at G0/G1 phase in renal cell carcinoma cells.

PLCε (phospholipase Cε), one of effectors belonging to the small GTPase superfamily, has been suggested to play a crucial role in carcinogenesis. However, its bio‐function in bladder cancer has never been demonstrated. In our previous study, we found that PLCε mRNA was highly expressed in bladder cancer tissues. In the present study, we silenced the PLCε gene by shRNA (small‐hairpin RNA) in the bladder cancer cell line BIU‐87. The results showed that it significantly inhibited cell proliferation and arrested the cell cycle at G0/G1‐phase. The regulation of cell characteristics has been related to PKCα (protein kinase Cα) activity. Further study showed that knockdown of the PLCε gene down‐regulated oncogenes c‐fos and c‐jun. These results indicate that PLCε plays a crucial role in bladder cancer, and PLCε may be a key molecule regulating the signal pathway of bladder cancer proliferation.

Objective To investigate the mechanism of vascular endothelial growth factor(VEGF) in influencing the angiogenesis of tumors through phospholipase C epsilon(PLCe)-NF-κB signal pathway.Methods Plasmid pGenesil-PLC for expression of PLC e-shRNA was transfected to human renal cell carcinoma 786-0 cell strain to silence the expression of PLCe gene.The expressions of PLC e,NF-κB and VEGF in transfected cells at mRNA level were determined by RT-PCR,while those at protein level by Western blot.The cells were treated with BAY11-7082,a specific inhibitor of NF-κB,and determined for inhibiting effect on cell proliferation by MTT method,then for expressions of VEGF mRNA and protein by RT-PCR and Western blot respectively.Results Recombinant plasmid pGenesil-PLCe inhibited the expressions of PLCe at mRNA and protein levels significantly,of which the inhibiting rates were 71.43% and 50.01% respectively.The plasmid also down-regulated the expressions of NF-κB and VEGF mRNAs and proteins significantly.BAY11-7082 showed significantly dose-and time-dependent inhibitory effect on the proliferation of 786-0 cells.Both the expressions of VEGF mRNA and protein in the cells after treatment with BAY11-7082 were inhibited significantly.Conclusion PLCe might inhibited the expression of NF-κB-dependent VEGF gene by inhibiting the expression of NF-κB gene and influence the angiogenesis of renal cell carcinoma.

探讨肝细胞黏附分子(hepatocyte cell adhesion molecule,hepaCAM)在肾癌786- 0细胞中不同表达水平对其凋亡及Caspase-3活性和表达的影响.将携带hepaCAM基因的重组质粒瞬时转染786- 0细胞,RT-PCR鉴定hepaCAM基因在786- 0中的表达;FCM检测细胞凋亡情况;分光光度计结合Western 印迹法检测hepaCAM表达与Caspase-3活性及蛋白水平表达的关系.结果显示,上调hepaCAM基因表达能诱导肾癌786- 0细胞凋亡,能上调Caspase-3活性及其蛋白水平且与hepaCAM表达水平呈正相关.上述结果表达,hepaCAM可能通过调节Caspase-3凋亡通路诱导肾癌786- 0细胞凋亡.

OBJECTIVES To evaluate hepaCAM (hepatocyte cell adhesion molecule) gene expression in patients with renal cell carcinoma (RCC) and to explore its effect on proliferation of 786-0 cells. hepaCAM is a tumor suppressor gene, which has been identified as a member of immunoglobulin superfamily cell adhesion molecule. METHODS Two-step reverse transcription- polymerase chain reaction was used to determine hepaCAM expression in 30 paired (RCC and the adjacent non-RCC) renal specimens. Transfection studies were carried out by expressing green fluorescent protein and green fluorescent protein-fused hepaCAM in 786-0 cells. RESULTS Significant downregulation of hepaCAM was detected in 25 of 30 RCC patients tested. When transfected into 786-0 cells, the number of colony formation was reduced by 5-fold according to colony formation assay. MTT (3-diphenyltetrazolium bromide) showed the inhibition rates on the fourth, fifth, and sixth days of culturing were 26.5%, 38.1%, and 35.7%, respectively. CONCLUSION Our data show that hepaCAM is frequently downregulated in RCC, and that exogenous hepaCAM exhibits antiproliferative effect on 786-0 cells, suggesting that silencing of hepaCAM may be associated with carcinogenesis of RCC. UROLOGY 75: 828-834, 2010. (C) 2010 Elsevier Inc.

Objective:To investigate whether PLCe gene down regulation by RNA interference(RNAi) leads to inhibition of proliferation in human bladder carcinoma T24 cell.Methods: The shRNA recombinant plasmids targeting to PLCe gene was constructed and transfected into bladder carcinoma T24 cell with Lipofectamine 2000.RT-PCR was used to monitor the validity of specific s h R N A in down regulation of PLCe.Then MTT assay was performed for detecting cell proliferation,the changes of PCNA were analyzed by immunocytochemical method,Electron microscope was used to observe the morphological changes.Results: The specific PLCe shRNA was confirmed to be efficient in silencing PLCe expression.PLCe gene down regulation by the shRNA recombinant plasmids inhibition cell proliferation rate about 78.01%,They were significantly different from that of control group(P0.01;After transfected the specific recombinant plasmids,PCNA expression was significantly decreased,They were significantly different from that of control group(P0.01;morphological changes produce apoptotic body.Conclusion: PLCeis likely to be potential molecular target for bladder carcinoma in gene therapy by RNAi.

Objective:To investigate the inhibitory effect of of protein kinase Cα(PKCα) activity by its specific inhibitor Safingol on bladder cancer cell proliferation.Methods:The bladder cancer BIU-87 cells were treated with PKCα specific inhibitor Safingol,PKCδ specific inhibitor Rottlerin,and PKC activator phorbol-12-myristate-13-acetate (PMA) ,respectively. Cell proliferation was determined by MTT assay. The expressions of c-fos and c-jun mRNA and protein were detected by RT-PCR and Western blotting,respectively. Cell cycle was detected by flow cytometry. Results:The proliferation of bladder cancer cells was significantly inhibited by PKCα specific inhibitor Safingol in a concentration-dependent,while PKCδ special inhibitor Rottlerin had no effects but could enhance the capability of cell proliferation. PKC activator PMA significantly enhanced the proliferation potency of BIU-87 cells. FCM analysis showed that inhibition of PKCα activity arrested cells at G0/G1 phase. RT-PCR and Western blotting showed that expressions of c-fos and c-jun were positively related with PKCα activity. Conclusion:Bladder cancer BIU-87 cell proliferation can be inhibited by PKCα specific inhibitor,which may be related to inhibition of c-fos and c-jun expressions.

Objective To construct a shRNA expression plasmid targeting phospholipase C epsilon(PLCε) and identify its function in bladder cancer cell line T24.Methods Potential RNAi oligonucleotides of PLCε were selected with the aid of online program.Then these oligonucleotides were synthesized and pGenesil-PLCε plasmids were constructed using pGenesil vector containing green fluorescent protein(GFP).The plasmids were digested by SalⅠ and sequenced to confirm the inserted sequence.RT-PCR and Western blot analysis were taken to detect the expression of PLCε at RNA and protein levels.The function of this PLCε RNAi in T24 cells was analyzed by MTT assay and flow cytometry.Results It was confirmed by digesting and sequencing that the 2 recombinant plasmids had been constructed successfully.The result of RT-PCR and Western blotting showed that pGenesil-PLCε plasmids inhibited PLCε expression in T24 cells obviously,and flow cytometry showed that the cell cycle was arrested at G0/G1 phase.MTT assay showed that proliferation of T24 cells was obviously inhibited.Conclusion pGenesil-PLCε effectively inhibits PLCε expression and cell proliferation in the bladder cancer cells.

Chronic cerebral hypoperfusion (CCH) increases the risk of Alzheimer disease (AD) through several biologically plausible pathways, but the relationship between CCH and the development of AD remains uncertain. To investigate expression of APP, BACE1 and Aβ in the hippocampus of BCCAO rats and study pathophysiological mechanism of AD from CCH. CCH rat model was established by chronic bilateral common carotid artery occlusion (BCCAO). Behavior was evaluated after BCCAO with Morris water maze and open-field task. Expression of Aβ was measured by enzyme linked immunosorbent assay (ELISA). β-Amyloid precursor protein cleavage enzyme 1 (BACE1) and β-amyloid precursor protein (APP) were tested by ELISA, Western blotting and RT-PCR. Cognitive impairment occurred with CCH by Morris water maze test and open-field task. The BACE1 and Aβ level in BCCAO rats was more increased than sham-operation control rats ( P < 0.01) but APP had no difference( P > 0.05). The expression of BACE1 and Aβ has no inter-grouop difference in BCCAO rats ( P > 0.05). The level of BACE1 and Aβ had positive correlation with cognitive impairment ( P < 0.01) while no correlation was observed between APP and cognitive impairment. Chronic cerebral ischemia contributes to cognitive impairment and vascular pathogenesis of Alzheimer’s disease that chronic cerebral hypoperfusion increases BACE1 and Aβ level in brain.

ObjectiveTo investigate serum levels of MMP-2, MMP-9, oxidized low-density lipoprotein (ox-LDL) in Alzheimer's disease (AD) patients and study the possible pathway and mechanism of AD with abnormal lipid metabolism.MethodsSubjects in this study were divided into 4 groups; normal lipid group without AD (N), hyperlipoidemia group without AD (H), normal group with AD (A), hyperlipoidemia group with AD (AH). There were 15 individuals in each group. MMP-2, MMP-9, ox-LDL was measured by enzyme linked immunosorbent assay (ELISA). Serum lipids levels were measured by biochemical methods.ResultsThe serum levels of MMP-2, MMP-9, ox-LDL were significantly higher in H. A and AH groups than those in N group. Those of ox-LDL in H, AH groups was higher than that of in A group. The serum level of MMP-2, MMP-9 in AH groups were higher than that of in H group. The score of mini-mental state examination (MMSE) in A and AD groups was negatively correlated with the serum level of ox-LDL. Relationship between the score of MMSE and the serum level of ox-LDL in AD groups and non-AD groups had statistical significance.ConclusionMMP-2, MMP-9, ox-LDL and abnormal lipid metabolism may participate in pathogenesis of AD, in which abnormal lipid metabolism induces expressions of MMP-2, MMP-9 and ox-LDL. Oxidative stress and blood-brain barrier disruption might accelerate the process of AD.