大鼠脂肪干细胞复合胶原-壳聚糖-硫酸软骨素三维支架构建组织工程软骨

Objective To evaluate the character of the collagen-chitosan-chondroitin sulfate scaffold seeded with rat adipose tissue-derived stromal cells. Methods A dipose tissue were harvested from 6 weeks old Wistar rats and the stromal cells were harvested by type Ⅰ collagenase and then cultured in vitro. Type Ⅰ collagen was fully mixed with chitosan, freeze-dried and cross-linked with chondroitin sulfate, then freeze-dried again and sterilized by ethylene oxide. The pore diameter, water content, porosity of the scaffold were tested. The adipose tissue-derived stromal cells were digested, seeded into the plates, scaffold, and cen-trifuged into pellet, and then induced into cartilage. MTT detection for cell proliferation was done. After 3 weeks, the cell morphology, and cell proliferation and adhesion were observed, and chondrngenic differenti-ation was also analyzed. Results The pore diameter, water content, porosity tested for the scaffold showed an appropriate form. Cell proliferation showed faster in the scaffold and pellet culture system after 5 day, there was still cell proliferation in the scaffold system after 14 days but no obvious changes in the pellet cul-ture system; ceils on the scaffold proliferated densely showed by histological staining, but there was a scaf-fold structure residues in the inner layer. The finding of type Ⅱ immunohistochemistry stain showed that cells express strong positive for type Ⅱ collagen in the scaffold and pellet culture system whereas it was weakly positive in the plate culture system; the specific mRNA for cartilage, type Ⅱ collagen, aggrecan and SOX-9 were expressed in all three systems showed by RT-PCR, but type X collagen was expressed continu-ously in the plate culture system and expressed after 21 days in the pellet culture system, whereas it was not detected in the collagen-chitosan-chondroitin sulfate scaffold system. Conclusion The parameters of the collagen-chitosan-chondroitin sulfate scaffold were suitable in our study. The results suggested that it can promote the adipose tissue-derived stromal cells proliferation and chondrogenic differentiation better than the plate and pellet culture systems and maintain the phenotype of chondrocytes well; it is the optimal choice for cartilage tissue engineering in the future.     

精子洗涤对精液中HBV DNA含量的影响

目的：检查HBV感染者精液是否存在乙型肝炎病毒（HBV），探讨精子洗涤是否能去除其中的HBV DNA。方法：用实时荧光定量PCR（FQ-PCR）法检测57名血清HBsAg阳性患者精液中HBV DNA，采用Percoll非连续梯度离心及上游法分离其中21份精液，并对洗涤后标本进行HBV DNA检测。结果：62份精液标本中有15份HBV DNA阳性，阳性率为24.19%;21份用于洗涤的精液标本中有，有7份HBV DNA阳性，洗涤后仍有6份标本阳性。结论：部分HBV感染者精液具有传染性，精子洗涤不能完全去除精液中的乙肝病毒。将HBV感染者的精液用于人工授精或体外受精应慎重。     

Cre recombinase-mediated site-specific recombination between plant chromosomes.

We report the use of the bacteriophage P1 Cre-loxsystem for generating conservative site-specific recombination between tobaccochromosomes. Two constructs, one containing a promoterless hygromycin-resistancegene preceded by a lox site (lox-hpt) and the other containing a cauliflowermosaic virus 35S promoter linked to a lox sequence and the cre coding region(35S-lox-cre), were introduced separately into tobacco plants. Crosses betweenplants harboring either construct produced plants with the two constructssituated on different chromosomes. Plants with recombination events wereidentified by selecting for hygromycin resistance, a phenotype expressed uponrecombination. Molecular analysis showed that these recombination eventsoccurred specifically at the lox sites and resulted in the reciprocal exchangeof flanking host DNA. Progenies of these plants showed 67-100% cotransmission ofthe new transgenes, 35S-lox-hpt and lox-cre, consistent with the preferentialcosegregation of translocated chromosomes. These results illustrate thatsite-specific recombination systems can be useful tools for the large-scalemanipulation of eukaryotic chromosomes in vivo.     

Asperger综合征与儿童孤独症的鉴别

对１５例Ａｓｐｅｒｇｅｒ综合征和３０例儿童孤独症的症状进行了对照分析，结果提示，Ａｓｐｅｒｇｅｒ综合征在语言交流、社会交往、运动等方面均存在不同于儿童孤独症的特点     

Influence of mild hypothermia on vascular endothelial growth factor and infarct volume in brain tissues after cerebral ischemia in rats

BACKGROUND: It has been demonstrated that mild hypothermia has obvious protective effect on both whole and local cerebral ischemia. However, the definite mechanism is still unclear for the brain protection of mild hypothermia on cerebral edema, inhibiting inflammatory reaction, stabilizing blood brain barrier, etc. OBJECTIVE: To investigate the effect of mild hypothermia on the expression of vascular endothelial growth factor and the infarct volume after cerebral ischemia in rats, and analyze the brain protective mechanism of mild hypothermia. DESIGN: A randomized grouping and controlled animal trial. SETTING: Department of Neurology, People's Hospital of Yunyang Medical College. MATERIALS: Twenty adult male SD rats of clean degree, weighing (250±30) g, were provided by the animal experimental center, School of Medicine, Wuhan University. The kits for SP immunohistochemistry were purchased from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. METHODS: The experiments were carried out in the laboratory of Department of Neurology, Renmen Hospital of Wuhan University from May to July 2005. ① The 20 rats were divided randomly into normal temperature group (n =10) and mild hypothermia group (n =10). Models of permanent middle cerebral artery occlusion were established with modified nylon suture embolization. The rats were assessed with the Longa standards: 0 point for without nerve dysfunction; 1 for mild neurological deficit (fore claws could no extend completely); 2 for moderate neurological deficit (circling towards the affected side); 3 for severe neurological deficit (tilting towards the affected side); 4 for coma and unconscious; 1-3 points represented that models were successfully established. The rats of the normal temperature group were fed at room temperature, and those in the mild hypothermia group were induced by hypothermia from 2 hours postoperatively, and the rectal temperature was kept at 34-35 ℃ for 72 hours. ② Measurement of infarct volume: All the rats were anesthetized by intraperitoneal injection overdose sodium pentobarbital 7 days postoperatively, and then the heads were cut down to harvest brain. The brain tissues were placed into -20 ℃ refrigerator for 20 minutes, coronal sections of 2 mm were prepared. The infarct sites were not stained, whereas normal brain tissues were stained as red. The infarct volumes were calculated by using MPLAS-500 multimedia color pathological image&&word analytical system. ③ Counting positive cells of vascular endothelial growth factor protein: The brains were harvested by cutting heads, then coronal sections of 2 mm were prepared. Routine dehydration, hyalinization, wax immersion and embedding were performed, then the detected with SP immunohistochemistry, the kits were purchased from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. The cells whose cytoplasm was yellow-brown were positive ones, a single sample as a unit, peri-ischemic site and ischemic core were selected, and the corresponding sites in controlateral hemisphere were taken as controls. Five visual fields were selected from each site to be observed under microscope, the cells were counted, and the average number of positive cells was calculated in each group. The numbers of positive cells were determined with the image analytical apparatus. MAIN OUTCOME MEASURES: Number of the positive cells of vascular endothelial growth factor protein; Infarct volume of rat brain tissue. RESULTS: All the 20 rats were involved in the analysis of results. ① Number of positive cells of vascular endothelial growth factor protein in brain tissue: It was obviously lower in the mild hypothermia group than in the normal temperature group [(24.02±5.05), (36.07±2.69) cells/high power visual field, P < 0.01]. ② Comparison of infarct volume of brain tissue: After MCAO, it was obviously smaller in the mild hypothermia group than in the normal temperature group [(153.25±23.14), (253.45±36.21) mm3, P < 0.01]. CONCLUSION: Mild hypothermia can inhibit the expression of vascular endothelial growth factor and decrease the volume of cerebral infarction. The inhibition of mild hypothermia on the expression of vascular endothelial growth factor may be one of the brain protective mechanisms.     

解释——护理过程中应重视的一个环节

在护理过程中护士和患者进行频繁接触和互动,护士主要应从以下几点做好解释工作,达到消除患者各种顾虑,改善和融洽医患关系,增强与患者的配合能力,使护理工作顺利进行,杜绝护理差错事故的发生,使患者心情愉快,早日康复.①解释环节对护士的素质要求,护士有高尚的职业道德水准,有扎实的基础知识和丰富的临床实践经验.②解释前,对患者的情况做全面了解.③整个护理操作前解释、操作中指导和操作后嘱咐的内容和要求.④鼓励患者及家属积极提出问题,并能全面、科学地做知识解答.笔者旨在通过撰写此文,引起护理同仁对解释环节的重视,不断提高解释水平的护理质量.     

缺氧诱导因子-1α在去势前后大鼠腹叶前列腺中表达的变化

目的探讨与缺氧相关的缺氧诱导因子-1α(HIF-1α)是否参与去势后前列腺萎缩过程.方法24只SD大鼠分为3组,其中A组(n=8)为假手术对照组,B组(n=8)为去势组,C组(n=8)为雄激素替代组(去势后肌注十一酸睾酮50mg/kg);术后3天处死,通过半定量RT-PCR检测与HIF-1α在去势前后前列腺表达变化.结果去势后大鼠前列腺的体积萎缩变小;雄激素替代组出现前列腺增生变大;对照组正常的大鼠前列腺有HIF-1 α mRNA低水平表达,去势组HIF-1α mRNA表达量增加,雄激素替代组HIF-1αmRNA表达量减少,与正常对照组比较,去势组的HIF-1α mRNA的表达量显著增加(P＜0.05),雄激素替代组的HIF-1αmRNA的表达量显著减少(P＜0.01).结论前列腺组织的缺氧参与去势后大鼠前列腺的早期萎缩过程.     

腹腔镜肾上腺肿瘤切除术75例报告

目的探讨腹腔镜下肾上腺肿瘤切除的方法和临床应用价值.方法1999年1月～2004年6月75例患者行腹腔镜肾上腺肿瘤切除术,其中采用经腹腔途径51例和经腹膜后途径24例.结果手术时间平均为(170.16±33.81)min,术中的出血量平均为(70.82±37.15)ml.术后平均住院时间为(5.87±1.01)d.中转开放手术2例(2.67%).发生并发症4例(5.33%),分别为膈肌损伤、胰腺损伤、肠道损伤、皮下血肿各1例.结论腹腔镜下行肾上腺肿瘤切除术具有创伤小,术中出血少,术后恢复快等优点,已经成为现代治疗肾上腺肿瘤的金标准.     

田径运动员最大摄氧量和无氧功测试中血乳酸的应用

目的:研究田径运动员最大摄氧量和Wingate测试后血乳酸的变化,分析血乳酸对测试结果的意义。方法:测试田径短跑运动员36例和中长跑运动员29例的最大摄氧量和Wingate无氧功,以及测试后的血乳酸值。结果:最大摄氧量测试后血乳酸数值为9～11mmol/L左右,摄氧量与乳酸值没有直接相关关系,但不同项目之间有一定的差异;Wingate测试后血乳酸数值一般大于12mmol/L,测试的主要评价指标与乳酸值都有较高的相关关系。结论:血乳酸值对于最大摄氧量测试意义不大,但对于Wingate测试可以作为一项辅助评价指标。