ReProComet: a new in vitro method to assess DNA damage in mammalian sperm.

The increasing request of chemical safety assessment demands for the validation of alternative methods to reduce the resort to animal experimentation. Methods that evaluate reproductive toxicity are among those requiring the largest use of animals. Presently, no validated in vitro alternative exists for the assessment of reproductive toxicity. Mammalian sperm are sensitive targets of DNA-reactive chemicals, which form premutagenic adducts. Here, we propose a new method based on comet assay to detect DNA damage induced by potential germ cell mutagens in bull sperm available from assisted reproduction practices. In somatic cells, chemical-induced adducts can be revealed by comet assay that detects DNA breaks produced during adduct repair. Mature sperm, however, are devoid of repair enzymes, and adducts are processed only after fertilization. For this reason, comet assay is not sensitive to detect DNA lesions induced in sperm by most chemicals. To overcome such limitation, we developed a modified comet assay based on the addition of a protein extract from HeLa cells to agarose-embedded sperm on microscopic slides. To test the method, sperm were treated in vitro with methyl methanesulfonate (MMS) or melphalan (MLP) and comet assay was conducted both with and without protein supplementation. No effect of MMS or MLP was detected without protein supplementation; on the contrary, a clear-cut dose-dependent effect was measured after addition of the cell extract. These results represent a proof of concept of a novel in vitro mutagenicity test on sperm that could offer a promising approach to complement previously validated in vivo germ cell genotoxicity assays.     

关于我国体外诊断试剂监督管理改革的建议

本文综述了美国、欧盟、日本、中国目前对体外诊断产品（试剂）监督管理的现状以及我国存在的问题。并根据我国具体情况提出了对体外诊断试剂监督管理的改革建议。     

Cleft closure for the treatment of unhealed perineal sinus

OBJECTIVE: Despite improvements in surgical practice, persistent perineal wound sinus is still a common complication after proctectomy. This study presents the success of a modified cleft closure technique in dealing with this problem. METHODS: From May 1997 patients with a persistent perineal sinus after surgery underwent a cleft closure - similar to that performed for patients with pilonidal sinus disease. RESULTS: Eight patients (6 male, 2 female) with an average age of 52 years underwent a cleft closure for a persistent perineal sinus after surgery. Four patients had undergone a proctocolectomy (ulcerative colitis), 2 an abdominoperineal excision of the rectum (adenocarcinoma) and 2 a proctectomy (1 Crohn's disease, 1 complication of diverticular disease). Symptoms had been present for an average of 41 months (range 5-152 months) and 3 patients had undergone other procedures attempted previously to deal with the problem. The first three patients had the procedure as an inpatient with an average stay of 4.7 days. The next 5 patients had the procedure as a day case (2 local anaesthetic, 3 general anaesthetic). Two patients developed a postoperative wound infection and all but one wound had healed completely by 8 weeks. In this patient the procedure was repeated to achieve healing. There was no other associated morbidity and no postoperative deaths. There have been no recurrences to date. CONCLUSION: Modified cleft closure for persistent perineal sinus is a simple procedure with low morbidity that can be performed under local anaesthetic in the day surgery unit.     

Association between cord blood IgE and genetic polymorphisms of interleukin-4, the β-subunit of the high-affinity receptor for IgE, lymphotoxin-α, and tumor Necrosis factor-α

High cord blood immunoglobulin E (cbIgE) is known to be associated with increased risks of atopic diseases in childhood. The relationship between genetic polymorphisms and high cbIgE has not been well documented. A cross-sectional study was conducted to assess the association between cbIgE and genetic polymorphisms of interleukin (IL)-4 -590C/T, the beta-subunit of the high-affinity receptor for IgE (FcepsilonRI-beta) E237G, lymphotoxin (LT)-alphaNcoI alleles, and tumor necrosis factor (TNF)-alpha -308G/A. A total of 320 mother-neonate pairs were recruited from four maternity hospitals from different locations of Taiwan. Cord blood was obtained and assayed for cbIgE. Polymerase chain reaction followed by restriction fragment length polymorphism was used to assess the genotypes. Three hundred pairs of mothers and neonates were included in the final analysis. Infants with IL-4 -590 C allele were found to have higher risk of elevated cbIgE (> or =0.35 IU/ml, 24.3%) (p = 0.004). After adjusting for gender, birth order, maternal age, and history of allergic disease in maternal and paternal families, odds ratios for CC and CT genotypes were 4.41 and 3.16 (95% confidence interval 0.78-22.67, and 1.66-6.13), respectively, using TT genotype as reference. The genotypes of FcepsilonRI-beta, LT-alpha, and TNF-alpha were not associated with cbIgE before or after the adjustment. Our finding suggested a significant association of cbIgE with genetic polymorphism of IL-4 -590C/T, but not with the genotypes of FcepsilonRI-beta, LT-alpha, and TNF-alpha.     

鼻内镜联合显微镜经鼻腔蝶窦及中颅窝手术

目的探讨鼻内镜下经鼻腔直接入路行蝶窦及中颅窝手术的方法. 方法鼻内镜下经较宽大一侧鼻腔将中鼻甲向外推移,鼻腔扩张器扩大视野,直达并开放蝶窦前壁, 鼻内镜与显微镜联合切除病变. 结果 10例孤立性蝶窦炎术后症状消失.6例蝶窦囊肿、脑膜瘤均一次手术切除.32例垂体腺瘤17例全切除,12例次全切除,3例大部分切除,术后补充X刀治疗.48例术后随访6个月～3.5年,平均2.5年,蝶窦囊肿、蝶窦炎、脑膜瘤无复发,3例垂体腺瘤复发,无颅内感染并发症,无鼻腔粘连、鼻出血等鼻腔并发症. 结论鼻内镜联合显微镜经鼻腔蝶窦及中颅窝手术损伤小、出血少、手术时间短、效果好.     

Improved Performance of Hip DXA Using a Novel Region of Interest in the Upper Part of the Femoral Neck: In Vitro Study Using Bone Strength as a Standard of Reference

We tested the hypothesis that bone mineral density (BMD) and bone mineral content (BMC) in proximal human femur specimens in the upper neck region of interest (ROI) and femoral neck axis length (FNAL) provide a significantly better prediction of femoral bone strength than standard ROIs in vitro. BMD and BMC were measured in 110 proximal femur specimens using a standard dual-energy X-ray absorptiometry (DXA) scanner. The analysis included a new ROI in the upper neck as well as the standard ROIs. FNAL was obtained from the scan images. The specimens' failure-load was measured in a mechanical loading device, simulating a fall on the greater trochanter. For the standard ROIs, correlations between failure-load and BMD ranged from R2 = 0.64 (shaft ROI) to R2 = 0.70, p < 0.001 (femoral neck). Prediction of strength by BMD did not significantly differ from those of BMC (R2 ranging from 0.65 to 0.75, p < 0.001). In the upper neck ROI, for both BMD and BMC correlations with failure-load were higher (R2 = 0.76 and 0.81, respectively; p < 0.001). A lower, yet still significant, correlation was found between FNAL and bone strength (R2 = 0.23, p < 0.001). Normalization of failure-load with respect to FNAL did not significantly increase the correlations with densitometric measures. This study provides in vitro evidence indicating that among the ROIs of the proximal femur the newly defined upper neck ROI provides the best prediction of bone strength. Only a weak association was observed between failure load and FNAL.     

老年人急性阑尾炎的腹腔镜手术治疗

目的 探讨老年人急性阑尾炎的临床特点，更好地把握腹腔镜阑尾切除（LA）术的方法和特点。方法 回顾分析50例老年人急性阑尾炎LA的临床资料。结果 LA均获成功，患者全部治愈，无严重的并发症发生，肠功能恢复时间及住院时间均缩短。结论 老年人反应迟钝，阑尾炎进展快，易发生坏疽及穿孔，合并症多，手术风险较高，LA是安全、可行的，除个别腹膜后阑尾炎外，其余均可施行LA。     

海绵窦上段动眼神经的应用解剖研究

目的：探讨海绵窦上段动眼神经的应用解剖并讨论其临床意义。方法：经血管灌注后观测10例尸颅的海绵窦上段动眼神经及其毗邻与血供。结果：海绵窦上段动眼神经与海绵窦上段背侧紧邻海马钩回，内侧与视交叉、后交通动脉毗邻，外上与脉络膜前动脉相邻，腹侧紧贴鞍背突。海绵窦上段动眼神经80％未见神经外营养血管，可见20％后交通动脉发出1～2根分支进入神经。结论：海绵窦上段动眼神经毗邻结构复杂，滋养血管细小；在海绵窦上缘手术时，应特别注意辨认和保护动眼神经。     

Reassessment of the accuracy of traditional sperm characteristics and adenosine triphosphate (ATP) in estimating the fertilizing potential of human semen in vivo

Receiver operating characteristic curves and accuracy parameters were computed for traditional sperm characteristics (concentration, motility, morphology) and the number of peroxidase negative cells, and the concentration of adenosine triphosphate (ATP) in semen from populations of fertile and infertile men, and men who achieved a pregnancy after varicocele treatment. The percentage and concentration per millilitre of spermatozoa with rapid linear progressive motility, and the ATP concentration, provided the best discrimination between fertile and treated fertile from infertile men. The misclassification rate was higher for sperm morphology, total progressive motility and viability, whereas sperm concentration and the total sperm count per ejaculate had the worst discriminating power. The number of peroxidase negative cells per 100 spermatozoa was highly specific in identifying men who achieved pregnancy after varicocele treatment. The lower limit of normality of sperm characteristics was remarkably different between fertile men and men achieving pregnancy after treatment or during infertility work-up.