2% Haemophilia A patients without mutation in the FVIII gene

In Germany, approximately 6,000 patients are suffering from haemophilia A. Screening methods cover 97% of the mutations. For the other patients the coding sequences of the FVIII gene have to be sequenced in total. Out of 1,350 patients, no mutation was observed in 80 patients. In 5 patients, we observed an inversion in intron 1. Known mutations were detected in 16 patients, and in 19 cases novel mutations were characterized (14 in coding regions and 5 in flanking introns). The mutations are mainly base pair substitutions, small deletions or insertions (max. 4 bp) and predicted to cause amino acid exchanges or frameshifts leading to premature stop codons. Moreover, 5 polymorphisms were identified in exons 14 and 26 as well as in introns 7 and 19. Further studies are necessary to identify their causative effects. Surprisingly, in 23 patients out of this subgroup of 80, no mutation was identified in the FVIII gene. Therefore, mutations in non-coding areas or even in other genes have to be considered responsible for the haemophilia A like phenotype. One of them codes for the von Willebrand factor (vWF). We confirmed in two of our cases mutations in the vWF gene.     

Aloe-emodin, an anthraquinone, in vitro inhibits proliferation and induces apoptosis in human colon carcinoma cells

The present study aimed to investigate the anticancer effect of aloe-emodin, an anthraquinone compound present in the leaves of Aloe vera, on two human colon carcinoma cell lines, DLD-1 and WiDr. Colon carcinoma cells were treated with various concentrations of aloe-emodin for different durations. Cell viability was measured by sodium 3'-[1-(phenylamino-carbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate assay. DNA fragmentation was analyzed by agarose gel electrophoresis. Nuclear shrinkage was visualized by Hoechst 33258 staining. Western blotting was used to indicate the release of apoptosis-inducing factor and cytochrome c from mitochondria and the phosphorylation of Bid. Caspase-3 and casein kinase II activities were measured by the respective assays. Cell viability analyses showed that aloe-emodin induced cell death in a dose- and time-dependent manner. Notably, the WiDr cells were more sensitive to aloe-emodin than the DLD-1 cells. Aloe-emodin caused the release of apoptosis-inducing factor and cytochrome c from mitochondria, followed by activation of caspase-3 leading to DNA fragmentation, nuclear shrinkage and apoptosis. In addition, exposure of colon carcinoma cells to aloe-emodin suppressed the casein kinase II activity in a time-dependent manner and was accompanied by a reduced phosphorylation of Bid, a downstream substrate of casein kinase II and a pro-apoptotic molecule. These findings showed that the inhibition of casein kinase II activity, the release of apoptosis-inducing factor and cytochrome c, and the caspase-3 activation are involved in aloe-emodin-mediated apoptosis in colon carcinoma cells.     

Two-year quality of life after breast cancer surgery: A comparison of three surgical procedures

Purpose

To analyze longitudinal changes in each subscale of a quality of life (QOL) measure and to explore their relationships to effective QOL predictors in breast cancer surgery patients.

Patients and methods

This prospective study analyzed 172 patients at two tertiary academic hospitals. All patients completed the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) and its supplementary breast cancer measure (QLQ-BR23) at baseline and at 1 and 2 years postoperatively. The 95% confidence intervals for differences in responsiveness estimates were derived by bootstrap estimation. Scores derived by these instruments were interpreted by generalized estimating equation (GEE) before and after surgery.

Results

A 2-year follow-up survey of the examined population revealed significant (P < 0.05) improvement in each QOL subscale. In both postoperative surveys, effect size was largest in the QLQ subscales for patients who had received mastectomy with reconstruction and lowest in those who had received modified radical mastectomy. After adjusting for time effects and baseline predictors, GEE approaches revealed the following explanatory variables for QOL: time, type of surgical procedure, age, chemotherapy, radiotherapy, hormone therapy, and preoperative functional status.

Conclusions

When evaluating QOL after breast cancer surgery, several factors other than the surgery itself should be considered. Patients should also be advised that their postoperative QOL might depend not only on the success of their operations, but also on their preoperative functional status.     

A phase II study of piritrexim in patients with advanced squamous head and neck cancer.

Piritrexim (PTX) is a newly developed lipid-soluble folate antagonist that crosses the cell membrane by a simple, rapid, carrier-independent diffusion process. A Phase II study was conducted to evaluate the activity of PTX in 34 patients with previously chemotherapy-naive squamous cell cancer of the head and neck area (SCCHN). Among them, 30 patients had received previous radiation therapy and/or surgery. Of 33 patients who could be examined, 3 had a complete response (CR), 6 had a partial response (PR), 11 had no change, and 13 had disease progression. The overall response rate (CR + PR) was 27% (9 of 33; 95% confidence interval, 13% to 46%). The response duration ranged from 36 to 360 + days (median, 162) and was similar to the best studies reported with methotrexate. The three most severe side effects (Grades 3 and 4 by World Health Organization criteria) were leukopenia, thrombocytopenia, and mucositis. These occurred in 41%, 26%, and 15% of the 34 patients, respectively. This study established PTX as an agent with some activity in SCCHN. The use of PTX in combination chemotherapeutic regimens needs to be explored.     

Feasibility of single-port laparoscopic cholecystectomy using a homemade laparoscopic port: a clinical report of 50 cases

Aim  

To report the clinical experience of transumbilical single-port laparoscopic cholecystectomy (TUSPLC), using a homemade laparoscopic access port composed of two inexpensive and common pieces of equipment readily available in the operating room.     

Two-port laparoscopic common bile duct exploration with T-tube choledochostomy for management of choledocholithiasis: an initial clinical report

Laparoscopic common bile duct exploration (LCBDE) is generally performed using a four- or five-port technique. We report a unique technique of two-port transcholedochal LCBDE with T-tube placement. Twelve consecutive patients with common bile duct (CBD) stones underwent LCBDE through two entry ports, one homemade single port (Uen port) inserted in a 2-cm umbilical wound and one 5-mm subxiphoid trocar port. With the assistance of a 1.2-mm needle that was inserted through a right lower intercostal space into the abdominal cavity to facilitate the operation, two-port dome-down laparoscopic cholecystectomy, choledochotomy, choledochoscopic removal of ductal caculi, and T-tube choldochostomy were performed with conventional methods using standard laparoscopic instruments along with manually operated angled shafts. After completion of the operation, the T-tube catheter was brought out through the subxiphoid trocar wound. All operations were completed successfully without the need of additional ports. There was no complication and no residual stones. Mean operation time was 120 minutes (range, 90 to 150 minutes), and mean postoperative hospital stay was 3.5 days (range, 3 to 4 days). Scarless wound healing was achieved except one T-tube scar. Two-port transumbilical LCBDE with T-tube choledochostomy is a feasible, safe, and effective technique that allows one-scar abdominal surgery for treatment of CBD stones. Further studies and the development of better instruments are necessary before this can be recommended as a standard procedure.     

Changing Approaches to Cholecystectomy in Elderly Patients: A 10-Year Retrospective Study in Taiwan

Background  

The prevalence of symptomatic gallbladder diseases increases with age. The present study evaluated cholecystectomy risk factors and hospital resource utilization in an elderly (aged 60 years and older) population of patients who had undergone open cholecystectomy (OC) or laparoscopic cholecystectomy (LC).     

Combretastatin A4-induced differential cytotoxicity and reduced metastatic ability by inhibition of AKT function in human gastric cancer cells

Combretastatin A4 (CA4) is a drug that targets tumor vasculature to inhibit angiogenesis. Whether CA4 has a direct effect on gastric cancer is not known. We herein investigated the effect of CA4 on growth and metastasis of gastric cancer cells at clinically achievable concentration and explored the associated antitumor mechanisms. Nine human gastric cancer cell lines, including two metastatic gastric cancer cell lines (AGS-GFPM1/2), constitutively expressing green fluorescence protein (GFP) were used. These metastatic AGS-GFPM1/2 cells expressed a higher level of phosphorylated serine 473 on AKT (p-AKT). Our results showed that CA4 (0.02-20 microM) has significant in vitro effects on reducing cell attachment, migration, invasiveness, as well as cell cycle G2/M disturbance on p-AKT-positive gastric cancer cells. In addition, a phosphoinositide 3-kinase inhibitor, LY294002 [2-(4-morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride], a specific AKT inhibitor, and 0.2 to 20 microM CA4 displayed a similar response profile on p-AKT-positive cells, suggesting that CA4-induced effect was mediated by inhibition of the PI3 kinase/AKT pathway. The results from in vivo GFP monitoring system indicated that CA4 phosphate (CA4-P; 200 mg/kg) significantly inhibited the s.c. and intra-abdominal growth of xenotransplanted AGS-GFPM2 cells in nude mice. Furthermore, CA4-P treatment showed a remarkable ability to inhibit gastric tumor metastasis as well as attenuate p-AKT expression. In conclusion, our study is the first to find that CA4 inhibited AKT activity in human gastric cancer cells. The decreased AKT activity correlated well with the CA4 antitumor growth response and decrease of metastasis. Further investigation on drugs targeting the PI3 kinase-AKT pathway may provide a new approach for the treatment of human gastric cancer.     

Post-cholecystectomy Quality of Life: A Prospective Multicenter Cohort Study of Its Associations with Preoperative Functional Status and Patient Demographics

Purpose  This study analyzed patient demographics and preoperative functional status for associations with post-cholecystectomy quality of life (QOL). Methods  This prospective study analyzed 159 cholecystectomy patients at two tertiary academic hospitals. All patients completed the SF-36 and the gastrointestinal quality of life index (GIQLI) at baseline and at 3 and 6 months postoperatively. The 95% confidence intervals for differences in responsiveness estimates were derived by bootstrap estimation. Scores derived by these instruments were interpreted by generalized estimating equation (GEE) before and after cholecystectomy. Results  The examined population significantly (p < 0.05) improved in both SF-36 subscales and GIQLI subscales. After adjusting for time effects (time, and time²) and baseline predictors, GEE approaches revealed the following explanatory variables for QOL: time, time², age, gender, preoperative GIQLI score, body mass index, and number of comorbidities. Conclusion  The data revealed dramatically improved post-cholecystectomy QOL. However, QOL change was simultaneously associated with preoperative functional status and demographic characteristics.