Development of Therapeutic dsP21-322 for Cancer Treatment.

Small activating RNAs (saRNAs) are a class of artificially designed short duplex RNAs targeted at the promoter of a particular gene to upregulate its expression via a mechanism known as RNA activation (RNAa) and hold great promise for treating a wide variety of diseases including those undruggable by conventional therapies. The therapeutic benefits of saRNAs have been demonstrated in a number of preclinical studies carried out in different disease models including cancer. With many tumor suppressor genes (TSGs) downregulated due to either epigenetic mechanisms or haploinsufficiency resulting from deletion/mutation, cancer is an ideal disease space for saRNA therapeutics which can restore the expression of TSGs via epigenetic reprogramming. The p21WAF1/CIP gene is a TSG frequently downregulated in cancer and an saRNA for p21WAF1/CIP known as dsP21-322 has been identified to be a sequence-specific p21WAF1/CIP activator in a number of cancer types. In this chapter, we review preclinical development of medicinal dsP21-322 for cancer, especially prostate cancer and bladder cancer, and highlight its potential for further clinical development.

Modulating the gut microbiota is involved in the effect of low-molecular-weight Glycyrrhiza polysaccharide on immune function.

Low molecular weight (6.5 kDa) Glycyrrhiza polysaccharide (GP) exhibits good immunomodulatory activity, however, the mechanism underlying GP-mediated regulation of immunity and gut microbiota remains unclear. In this study, we aimed to reveal the mechanisms underlying GP-mediated regulation of immunity and gut microbiota using cyclophosphamide (CTX)-induced immunosuppressed and intestinal mucosal injury models. GP reversed CTX-induced intestinal structural damage and increased the number of goblet cells, CD4+, CD8+ T lymphocytes, and mucin content, particularly by maintaining the balance of helper T lymphocyte 1/helper T lymphocyte 2 (Th1/Th2). Moreover, GP alleviated immunosuppression by down-regulating extracellular regulated protein kinases/p38/nuclear factor kappa-Bp50 pathways and increasing short-chain fatty acids level and secretion of cytokines, including interferon-γ, interleukin (IL)-4, IL-2, IL-10, IL-22, and transforming growth factor-ß3 and immunoglobulin (Ig) M, IgG and secretory immunoglobulin A. GP treatment increased the total species and diversity of the gut microbiota. Microbiota analysis showed that GP promoted the proliferation of beneficial bacteria, including Muribaculaceae_unclassified, Alistipes, Lachnospiraceae_NK4A136_group, Ligilactobacillus, and Clostridia_vadinBB60_group, and reduced the abundance of Proteobacteria and CTX-derived bacteria (Clostridiales_unclassified, Candidatus_Arthromitus, Firmicutes_unclassified, and Clostridium). The studies of fecal microbiota transplantation and the pseudo-aseptic model conformed that the gut microbiota is crucial in GP-mediated immunity regulation. GP shows great potential as an immune enhancer and a natural medicine for treating intestinal inflammatory diseases.

Flank-suspended versus prone percutaneous nephrolithotomy: changes of haemodynamics, arterial blood gases and subjective feelings.

BACKGROUND: The flank-suspended position was adopted in percutaneous nephrolithotomy (PCNL), and haemodynamics, blood gas variables and subjective feelings were examined with an attempt to explore the effect of the operative position in PCNL on the body's inner environment and patient comfort. OBJECTIVE: The influence of the flank-suspended and prone position on haemodynamics, arterial blood gases and subjective feelings in patients receiving PCNL was examined. DESIGN, SETTING AND PARTICIPANTS: A total of 100 patients with kidney stones who underwent PCNL during January 2010 to January 2011 were divided into flank-suspended groups (n = 50) and prone groups (n = 50) at random in terms of the operative position. The blood pressure, heart rate, respiratory frequency and oxyhaemoglobin saturation and blood gas variables were determined at different time points (before the operation, after position change, 30 min after the start the operation and immediately after the operation). Visual analogue scale (VAS) scoring system was employed to define the posture comfort, dyspnoea and puncture-site pain 24 h postoperation in the patients. All the measures were compared between patients in different positions at different time points. STATISTICAL ANALYSIS: Paired t-test was employed in the comparison of measures detected at different time points in the same group. Two-group comparison was subjected to t-test. A p value less than 0.05 was considered statistically significant. RESULTS AND LIMITATIONS: The blood pressure was decreased within and after the operation in both groups, substantially in the prone group, significantly lower than that before the operation (p<0.05). No significant differences in the heart rate, respiratory frequency and oxyhaemoglobin saturation were noted among the different time points in the same group. Blood gas analysis showed that pH value and base excess were profoundly reduced within and after the operation in the two groups, significantly lower than those before the operation, and the decrease was most manifest in the prone group. There was no difference in the blood sodium and potassium among the different time points in each group. The flank-suspended group was superior to the prone group with regard to posture comfort and dyspnoea degree but not puncture-site pain 24 h postoperation. CONCLUSIONS: Flank-suspended and prone PCNL affects the haemodynamics, blood gas variables and subjective feelings of patients to a varying degree. The flank-suspended PCNL possesses advantages over prone PCNL such as little influence on haemodynamics and blood gas variables, satisfactory posture comfort, less dyspnoea and easy access to vital sign observation.

Regulatory Variants and Disease: The E-Cadherin -160C/A SNP as an Example.

Single nucleotide polymorphisms (SNPs) occurring in noncoding sequences have largely been ignored in genome-wide association studies (GWAS). Yet, amounting evidence suggests that many noncoding SNPs especially those that are in the vicinity of protein coding genes play important roles in shaping chromatin structure and regulate gene expression and, as such, are implicated in a wide variety of diseases. One of such regulatory SNPs (rSNPs) is the E-cadherin (CDH1) promoter -160C/A SNP (rs16260) which is known to affect E-cadherin promoter transcription by displacing transcription factor binding and has been extensively scrutinized for its association with several diseases especially malignancies. Findings from studying this SNP highlight important clinical relevance of rSNPs and justify their inclusion in future GWAS to identify novel disease causing SNPs.

Flank suspended supine position for percutaneous nephrolithotomy.

AIMS: Prone and supine positions for percutaneous nephrolithotomy are widely used but have their drawbacks. We report a new positioning method called "flank suspended supine position" (FSSP) for PCNL and describe our experience with PCNL in this position to evaluate its safety and efficacy. METHODS: Retrospective study of 150 cases of renal stone patients treated with PCNL in a new position called flank suspended supine position (FSSP) from June 2009 to July 2010. All patients were treated with PCNL in FSSP under epidural anesthesia. Operation time, bleeding rate, stone free rate, and complications were recorded. RESULTS: All patients tolerated FSSP. Mean operation time was 78.29±26.13 min. Initial stone-free rate was 83%. For those with residual stones (26 cases), 18 were stone-free after a second PCNL, 8 after extracorporeal shock wave lithotripsy (ESWL). Mean hospital stay was 7.63±2.39 days. No penetrating injury of the pleural cavity or injury to visceral organs was reported. SUMMARY: FSSP is an effective and safe position for PCNL in our hands and its effectiveness relative to traditional prone position needs to be determined in future randomized studies.

Histone deacetylase inhibitor trichostatin A inhibits the growth of bladder cancer cells through induction of p21WAF1 and G1 cell cycle arrest.

PURPOSE: To investigate whether Trichostatin A (TSA) possesses antitumor activity against human bladder cancer cells, and if any, its mechanism. MATERIALS AND METHODS: A human bladder cancer cell line, BIU-87, was treated with different concentrations of TSA. After treatment, cell growth was measured by MTT assay. Cell apoptosis and cell cycle changes were examined by means of flow cytometry (FCM). Apoptosis was confirmed by apoptotic ladder formation assay. mRNA expression of p21WAF1 and p53 was assessed by differential reverse transcription-polymerase chain reaction. RESULTS: Trichostatin A significantly inhibited the proliferation of bladder cancer cell at nanomolar concentrations in a time- and dose-dependent fashion. TSA treatment caused cell cycle arrest at the G1 phase and increased apoptotic cell death as shown by FCM and DNA fragmentation analysis, accompanied by increased p21WAF1 mRNA expression. In addition, TSA treatment did not alter p53 mRNA expression. CONCLUSION: Our results indicate that TSA is able to inhibit bladder cancer cell growth in vitro, possibly through p21WAF1 mediated cell cycle arrest and apoptotic cell death. This study suggests that TSA may be a potential therapeutic agent for the treatment of bladder cancer.