Preferential loss of Fhit expression in signet-ring cell and Krukenberg subtypes of gastric cancer

Gastric cancer of youth is predominantly a disease of women, usually of the signet-ring cell subtype, with a predilection for metastasizing to the ovaries. The metastatic ovarian tumor is named a Krukenberg tumor. However, the characteristic genetic alterations between the primary gastric cancer and its metastatic ovarian tumor have not been studied. We used laser capture microdissection to procure tissues from 7 patients with gastric cancer who had ovarian metastases (Krukenberg tumor) and tissues from 14 patients with gastric cancer without ovarian metastases. Loss of heterozygosity (LOH) analysis was performed by use of 16 polymorphic markers, which are mapped to the FHIT, APC, p16, BRCA2, E-cadherin, p53, BRCA1, and DPC4 loci. Immunohistochemical staining with anti-Fhit antibody was performed in 7 Krukenberg tumors and 92 gastric cancers without ovarian metastases. LOH at the FHIT locus was observed in six (85.7%) of the seven Krukenberg tumors. In contrast, the gastric cancers without ovarian metastases showed a lower frequency (28.6%, 4/14) of LOH at the FHIT locus (p < 0.05, odds ratio = 1/15). Anti-Fhit antibody showed that expression of Fhit was lost in each of the 7 (100%) Krukenberg tumors but in only 41 (44.6%) of the 92 patients who had gastric cancer without ovarian metastases (p < 0.05; odds ratio = 1/18.614). Further analysis showed that loss of Fhit expression is highly associated with signet-ring cell type gastric cancer (p < 0.0001, odds ratio = 62.5) but is not correlated with prognosis. Alteration of the FHIT gene is a characteristic of signet-ring cell type gastric cancer and Krukenberg tumor.     

L-精氨酸对大鼠蛛网膜下腔出血脑组织血供的改善作用

目的　探讨L 精氨酸 (L Arg)对蛛网膜下腔出血(SAH)后继发性脑缺血的影响。方法　用血管内脑底动脉环穿刺法建立大鼠SAH模型 ,将动物分为假手术 (SO)组、SAH组和SAH +L Arg组。L Arg于术前 3 0min按 50 0mg·kg- 1ip ,每 6h追加 1次。动态检测 2 4h内大脑顶叶皮层局部脑血流量 (rCBF) ,测定不同时间点血清一氧化氮 (NO ,NO-2 /NO-3 )水平 ,同时监测血压和血气。结果　假手术对rCBF和血清NO含量无显著影响。SAH组术后rCBF迅速降低并持续 2 4h ;SAH组血清NO水平于术后 1h开始下降 ,持续 2 4h ;与SAH组比较 ,SAH +L Arg组rCBF下降的速度减慢且程度减轻 ;血清NO水平下降的程度减轻。结论　L Arg可部分补偿SAH后血清NO不足 ,从而缓解继发性脑缺血程度     

Quiescent nasal T/NK cell lymphoma manifested as primary central nervous system lymphoma

A 57-year-old man was diagnosed as primary T/NK-cell central nervous system lymphoma (CNSL) with intraocular involvement. However, review of a surgical specimen taken three years before for chronic paranasal sinusitis revealed an overlooked nasal T/NK cell lymphoma (TNKL), which showed similar histomorphology and immunophenotype with the CNS disease. Another patient, a 43-year-old woman, was initially diagnosed as a rare primary leptomeningeal T-cell lymphoma with ocular manifestation. Three years later, an isolated nasal TNKL emerged. Immunohistochemical and cytogenetic studies confirmed the same nature of the CNSL and the nasal TNKL. The nasal TNKLs of both patients had a strong expression of CD3, CD56, and Epstein-Barr virus antigens, but features of angiodestruction and mucosal ulceration were absent. We propose that: 1. a locally silent “quiescent” form of nasal TNKL may exist; and 2. a thorough examination and even blind biopsy of the nasal cavity is indicated when primary T/NK-cell CNSL is diagnosed. Am. J. Hematol. 60:161–163, 1999. © 1999 Wiley-Liss, Inc.     

卵裂期发育延缓胚来源的人类胚胎干细胞系的建立

目的观察体外受精（IVF）周期中24h内不发生卵裂的第3天（D3）发育延缓胚的体外发育潜能,为建立人胚胎干细胞库提供新的材料来源。方法通过囊胚序贯培养法将卵裂期发育延缓胚培养至囊胚,观察囊胚形成率及囊胚质量。用逐步logistic回归分析囊胚形成率与卵裂球数、卵裂球对称性及卵裂球碎片比率的相关性。用免疫外科法去除滋养细胞,将得到的原代克隆接种于小鼠胚胎成纤维细胞上,体外传代并鉴定。结果120枚卵裂期发育延缓胚培养出囊胚22枚（囊胚形成率18．7％）。第6天早期囊胚、囊胚、满囊胚、扩张胚、孵化胚及孵化后囊胚的比率分别为5．9％、23．5％、35．3％、23．5％、5．9％及5．9％。内细胞团分级及滋养外胚层分级均仅有1个为A级,其余均为B或C级。发育延缓胚的囊胚形成率与卵裂球数、卵裂球对称性相关,而与卵裂球碎片比率无关。获得内细胞团7个,原代克隆3个,成功培养出2株人类胚胎干细胞系（hESC）,并具有hESC的生物学特性：碱性磷酸酶（AKP）、阶段特异性胚胎抗原（SSEA）4、肿瘤排斥抗原（TRA）1-60、TRA-1-81呈阳性;悬浮培养可形成拟胚体;在严重免疫缺陷（SCID）小鼠体内可形成畸胎瘤;分别传45及35代后核型正常。结论部分发育延缓胚可以发育为囊胚,从而可为建立人胚胎干细胞系提供新的材料来源。     

自发性脑出血早期血肿增大

脑出血早期血肿增大严重影响患者的预后。文章对脑出血早期血肿增大的发生率、诊断标准、影响因素和防治方法进行了综述。     

Foxp3在神经母细胞瘤LA-N-6细胞的表达及化疗药物对其影响

目的：研究神经母细胞瘤细胞是否表达免疫逃逸相关分子Foxp3及其对化疗药物的敏感性。方法：体外培养神经母细胞瘤细胞株LA-N-6;MTT试验明确化疗药物环磷酰胺（CTX）、盐酸吡柔比星(THP)对LA-N-6的效应剂量,流式细胞术(FACS)及实时定量PCR（RT-PCR）测定Foxp3在LA-N-6中的表达和CTX、THP对LA-N-6中Foxp3表达的影响。结果：FACS结果显示LA-N-6细胞高表达Foxp3,加入 CTX、THP后显著降低了LA-N-6上Foxp3的表达（P<0.05）;RT-PCR显示CTX显著降低了LA-N-6细胞中Foxp3的表达（P<0.05）,THP加药组与对照组比较差异无统计学意义。结论：肿瘤逃逸相关转录因子Foxp3高表达于神经母细胞瘤细胞LA-N-6;CTX、THP能分别在蛋白或基因水平降低Foxp3分子在LA-N-6的表达,提示CTX、THP可能通过抑制Foxp3分子的表达而抗肿瘤的作用。 ［中国当代儿科杂志,2010,12（5）：386-389］     

细菌脂蛋白耐受对脓毒症老龄及成年小鼠心脏功能保护作用的对比

目的 探讨细菌脂蛋白（BLP）耐受对老龄小鼠心肌细胞的保护作用,并比较其对老龄小鼠与成年小鼠心功能影响的差异。方法 选取86只健康雄性C57BL/6老龄小鼠（SPF级,24月龄）和55只成年雄性C57BL/6小鼠（6～8周龄）,用小剂量BLP预处理老龄小鼠及成年小鼠诱导BLP耐受,分别比较正常对照组、假手术（sham）组、脓毒症（剖腹手术行盲肠结扎穿孔致脓毒症,CLP）组、BLP耐受＋CLP组,其中成年小鼠在盲肠结扎穿孔0,2,6,12h四个时间点做小鼠二维超声心动图,老龄小鼠在0,3,6,12h四个时间点做小鼠二维超声心动图,选择左心室短轴缩短率（FS）、射血分数（EF）、左心室舒张末期内径（LVIDd）等做为观测指标。结果 老龄小鼠CLP组与BLP耐受＋CLP组的FS、EF、LVIDd自手术后均呈下降趋势,但是BLP耐受＋CLP组的降幅较CLP组小。成年小鼠BLP耐受＋CLP组0～6h时段FS、EF、LVIDd的变化趋势与CLP组相同,但各时间点数值均低于CLP组;而6h后BLP耐受＋CLP组的3项指标转而上升,至12h时高于CLP组,其中FS、EF差异均有统计学意义（P＜0.05）。BLP耐受＋CLP组老龄小鼠的FS呈现出进行性下降趋势,而在成年小鼠中6h后为进行性增高趋势,12h明显高于假手术组和CLP组（P＜0.05）。成年小鼠CLP组在术后EF上升,6h后出现下降的趋势,12h低于术前水平;而老龄小鼠CLP组术后EF进行性下降。成年小鼠BLP耐受＋CLP组术后出现的EF值呈逐渐上升趋势,相反,老龄组则进行性下降。老龄小鼠BLP耐受+CLP组的LVIDd术后进行性下降,而成年小鼠BLP耐受＋CLP组术后2h时低于术前水平（P＜0.05）,6h后逐渐回升。结论 脓毒症对FS、EF、LVIDd有抑制作用,老龄小鼠心脏收缩功能和舒张功能所受抑制更加严重;BLP耐受对心功能有一定的保护作用,此作用在成年小鼠较明显,在老龄小鼠未见明显保护作用。     

Effects of blockade of cerebral lymphatic drainage on regional cerebral blood flow and brain edema after subarachnoid hemorrhage

The study was designed to observe the influence of blockade of cerebral lymphatic drainage on the regional cerebral blood flow (rCBF) and brain edema after experimental subarachnoid hemorrhage (SAH). Wistar rats were divided into non-SAH, SAH, and SAH plus cervical lymphatic blockade (SAH + CLB) groups. Autologous arterial hemolysate was injected into rat's cisterna magna to induce SAH. The rCBF was recorded continuously by a laser Doppler flowmeter. Intracranial pressure (ICP) was also monitored. After 24 hours and 72 hours of SAH, the rats were sacrificed and the brain was harvested for water content detection. It was found that there was no obvious change of rCBF and brain water content during the experiment in non-SAH group. An immediate and persistent drop in rCBF was found in SAH group. The drop in rCBF was more obvious in SAH + CLB group. CLB also worsened the SAH-induced increase in ICP. The brain water content 24 hours and 72 hours after induction of SAH in SAH group increased significantly. CLB led to a further increase of brain water content. In conclusion, blockade of cerebral lymphatic drainage pathway deteriorates the secondary cerebral ischemia and brain edema after SAH.     

Expression of the receptors of VEGF and the influence of extract of Ginkgo biloba after cisternal injection of autologus arterial hemolysate in rats

The study was aimed to investigate the alterations of vascular endothelial growth factor (VEGF) receptors and the influence of extract of Ginkgo biloba (EGb) after subarachnoid hemorrhage (SAH). Wistar rats were divided into non-SAH, SAH, vehicle, EGb1 (lower dose), and EGb2 (higher dose) groups. Autologus arterial hemolysate was injected into cisterna magna to induce SAH. The non-SAH rats received cisternal injection of saline instead. Rats underwent RT-PCR determination of one of the VEGF receptors flt-1mRNA, and immunohistochemistry for VEGF receptors Flt-1 and Flk-1. The results revealed that there was only slight expression of flt-1mRNA in the brain tissue in non-SAH rats. The expression in SAH group was enhanced 24 hours and 72 hours after cisternal injection. No Flt-1 and Flk-1 positive cell was observed in the brain in non-SAH group. A good few Flt-1 and Flk-1 positive cells were found in cortex and other regions of the brain in SAH group. The expression of flt-1mRNA, Flt-1 and Flk-1 proteins were increased by the use of two doses of EGb. It was concluded that the up-regulated expression of the two kinds of VEGF receptors may be an intrinsic protective mechanism in the process of SAH, which can be enhanced by EGb.     

抗肌萎缩蛋白小基因对Duchenne肌营养不良病理改变的治疗作用

目的　探讨Duchenne肌营养不良 (DMD)的病理变化和治疗方法。方法　采用重组腺相关病毒载体 (rAAV)介导的人抗肌萎缩蛋白小基因 (SMCKA3999)观察DMD的病理改变和治疗作用。将抗肌萎缩蛋白小基因SMCKA3999克隆至rAAV并包装成rAAVSMCKA3999病毒 ,将 5× 10 9病毒颗粒分多点注射于DMD模型鼠mdx腓肠肌 ,4个月后取局部肌组织 ,分别采用免疫荧光、埃文斯氏蓝染料、电镜等方法 ,观察rAAVSMCKA3999对DMD病理改变的治疗作用。结果　rAAVSMCKA3999有效稳定表达并使肌膜缺失的抗肌萎缩蛋白恢复 ,明显改善DMD肌肉病理改变。结论　rAAVSMCKA3999能显著改善mdx小鼠骨骼肌病理变化 ,有希望成为Duchenne肌营养不良有效的生物治疗药物