Deletion of aldose reductase leads to protection against cerebral ischemic injury.

Previously, we reported that transgenic mice overexpressing endothelin-1 in astrocytes showed more severe neurological deficits and increased infarct after transient focal ischemia. In those studies, we also observed increased level of aldose reductase (AR), the first and rate-limiting enzyme of the polyol pathway, which has been implicated in osmotic and oxidative stress. To further understand the involvement of the polyol pathway, the mice with deletion of enzymes in the polyol pathway, AR, and sorbitol dehydrogenase (SD), which is the second enzyme in this pathway, were challenged with similar cerebral ischemic injury. Deletion of AR-protected animals from severe neurological deficits and large infarct, whereas similar protection was not observed in mice with SD deficiency. Most interestingly, AR(-/-) brains showed lowered expression of transferrin and transferrin receptor with less iron deposition and nitrotyrosine accumulation. The protection against oxidative stress in AR(-/-) brain was also associated with less poly(adenosine diphosphate-ribose) polymerase (PARP) and caspase-3 activation. Pharmacological inhibition of AR by Fidarestat also protected animals against cerebral ischemic injury. These findings are the first to show that AR contributes to iron- and transferrin-related oxidative stress associated with cerebral ischemic injury, suggesting that inhibition of AR but not SD may have therapeutic potential against cerebral ischemic injury.     

Inhibition of post-meningitic cochlear injury with cerebrospinal fluid irrigation.

OBJECTIVE: Labyrinthitis ossificans, the pathologic ossification of the otic capsule associated with profound deafness and loss of vestibular function occurs frequently as a sequella of bacterial meningitis and subsequent purulent labyrinthitis. Experimentally, in Streptococcus pneumoniae meningitis, it has been shown that a vigorous inflammatory response to teichoic acids in the bacterial cell wall contributes to cochlear damage and subsequent fibrosis and ossification. The hypothesis of this study is that a dilution of concentration of inflammatory mediators through cerebrospinal fluid (CSF) irrigation will lead to a reduction in both inner ear pathology and permanent hearing loss. STUDY DESIGN AND SETTING: Auditory brainstem response testing was used to determine baseline hearing thresholds in 20 Mongolian gerbils (12 irrigated, 8 sham irrigated animals) at 32 kHz, 16 kHz, 8 kHz, and 4 kHz frequencies. Their thresholds at 14 days and 120 days post-procedure were also obtained. Streptococcus pneumoniae meningitis was induced in both groups of animals by intrathecal (i.t.) injection of bacteria. Both groups received penicillin treatment. Forty-eight hours after inoculation, both groups were implanted with i.t. inflow and outflow catheters. The irrigated group was infused continuously with artificial CSF over 36 hr at a rate of 70 muL/hr and the outflow sampled. The tubing in the sham irrigated group was clamped (without sampling). They were sacrificed at 120 days post-procedure and histomorphometric analysis carried out. The concentration of interleukin 1beta (IL-1beta) for the CSF samples from the irrigated group were compared to samples collected from an additional control group of 8 non-irrigated meningitic gerbils. IL-1beta was chosen to study because it is a potent pro-inflammatory cytokines in bacterial meningitis that is unaffected by the neurosurgical trauma of the experimental protocol. RESULTS: Twenty animals survived the meningitis (6 irrigation, 6 sham irrigation, 8 non-irrigation meningitic controls). At Days 14 and 120 post-infection, the irrigated animals manifested significantly less hearing loss with a mean loss of 28.82 dB compared to the sham irrigation group mean loss of 40.76 dB (P < 0.03). The degree of hearing loss in both groups was frequency-dependent with greater loss at higher frequencies (mean loss = 22.4 dB at 32 kHz, 23.0 dB at 16 kHz, 18.6 dB at 8 kHz, and 12.5 dB at 4 kHz). Histomorphometric analysis demonstrated a marked reduction in degeneration of the spiral ligament, spiral ganglion cells, and stria vascularis in experimental animals as compared to controls. Immunohistochemistry showed a significant reduction in IL-beta1 concentrations in the irrigated animals compared to the non-irrigated, infected controls (P < 0.03). CONCLUSIONS: Irrigation of CSF resulted in a significant reduction in post-meningitic cochlear injury when compared to controls. This model for continuous cerebrospinal fluid irrigation provides a means to evaluate the effects of a dilution of inflammatory mediators on hearing loss and labyrinthitis ossificans after bacterial meningitis. SIGNIFICANCE: Despite advances in the prevention of meningitis and improved antibiotic treatment, bacterial meningitis continues to have significant associated morbidity. This study provides insight into some of the mechanisms responsible for post-meningitic hearing loss and labyrinthitis ossificans and presents a novel approach to reduce these complications.     

Buccal mucosal graft for urethral reconstruction.

Urethral reconstruction with graft substances, such as skin and bladder mucosa, has been previously used when primary anastomosis cannot be achieved. However, stricture and meatal prolapse are associated with these grafts. We report the use of buccal mucosa for the reconstruction of urethral defects in 3 patients. One patient with failed operation for hypospadias received tube buccal mucosal graft for urethral replacement. Two patients with urethral necrosis and stricture received onlay buccal mucosal graft. All patients were disease-free during follow-up (range, 12 to 49 months; mean, 36 months). One patient had a pinhole fistula that was successfully managed with simple repair. This technique appears to be useful for urethral reconstruction when a local graft is not available, even in patients with complicated conditions.     

赖型钩体flaB2与VR1012中的CpG基序分析

目的：对问号赖型钩端螺旋体（赖型钩体）DNA疫苗[包括内鞭毛蛋白基因（flaB2)和质粒DNA表达载体（VR1012）]的CpG基序（CpG motifs)进行分析，为DNA疫苗免疫机制的阐明和提高DNA疫苗的效能奠定基础。方法：以flaB2与VR1012构建重组DNA的免疫原，对flaB2及VR1012全核苷酸序列进行计算机分析（分类、计数和定位）。结果：CpG的“C”的侧翼为两个嘌呤，“G”的侧翼为两个嘧啶，在flaB2中共3个，分别为GACGCT，GACGTC和GACGCC；在VR1012中共11个，分别为GACGTC1个，GACGCT2个，GACGCC1个，GACGTT1个，GGCGTT2个，GGCGCT2个，GGCGCC1个，AACGCT1个，其中特别重要的TGACGTCA4个和TAACGCCA有1个，位于5'端456－463；509－516；592－599；778－785和486－493；4个TGACGTCA和1个TAACGCCA均位于5'端且相对集中。结论：赖型钩体flaB2与VR1012构成的DNA疫苗含有TGACGTCA等CpG，这些基序又称免疫刺激序列，构成了DNA疫苗中的佐剂。     

End-to-end oesophageal anastomosis: an experimental study in the rat

To study the effects of suture material, calibre and tension on the end-to-end oesophageal anastomosis, the cervical oesophagus of the rat was resected for 2.5 mm, 7.5 mm and 12.5 mm respectively in three groups of 60 rats each. End-to-end anastomosis was performed with either 6/0 silk, 6/0 polypropylene or 9/0 polypropylene sutures. There was no postoperative leakage. Mortality was mainly due to hair bolus obstruction at the site of anastomosis. There was no difference in mortality and anastomotic stenosis between groups with different resection lengths. Within each group, however, mortality and anastomotic stenosis were significantly higher in the subgroup of rats with 6/0 silk suture than that in the subgroup with 6/0 polypropylene sutures, which also had significantly higher mortality and anastomotic stenosis than the subgroup with 9/0 polypropylene sutures. Histological examination of the stenosed anastomosis showed disruption of muscle layers and submucosal thickening. It is concluded that the use of fine calibre sutures of biologically inert material would be preferred for the end-to-end anastomosis of the oesophagus.