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.     

SEN病毒D亚型ORF1-C端基因的克隆、表达和鉴定

目的:获得SENV-D亚型ORF1-C端蛋白基因序列,并进行表达,为进一步研究及诊断、治疗应用奠定基础.方法:用PCR法从SENV阳性血清中获得SENV-DORF1C端基因,测序验证后,构建了pQE30-SENV-D重组表达质粒,并经过转化E.coli M15,IPTG诱导表达,Western blot分析表达结果.结果:获得了正确序列的SENV-D亚型ORF1-C端蛋白基因序列,表达出Mr约为38×103的蛋白.表达蛋白能与患者血清中相应的抗体结合,发生抗原抗体反应.结论:成功获得并表达了SENV-D-ORF1-C端蛋白,并经Western blot印迹法证实能与阳性血清中的抗体发生抗原抗体反应,有可能用于检测SENV-D抗体.为今后进一步研究有助于疫情监测及早期发现感染者的抗体奠定了坚实的基础.     

C型臂X线机引导下经皮双侧骶髂拉力螺钉固定治疗骶骨H形骨折

目的探讨骶骨H形骨折可行的治疗方法。方法运用C型臂X线机引导下经皮双侧骶髂拉力螺钉固定治疗骶骨H形骨折15例。结果15例患者均获随访，随访时间7—34个月，骨折临床愈合时间3～5个月，术后均未留下明显行走障碍，下蹲等活动接近正常。结论在C型臂X线机精确引导下，经皮双侧骶髂拉力螺钉固定技术能有效地固定骶骨H形骨折中的垂直骨折，纠正骨盆垂直方向移位，操作简洁安全，疗效可靠。     

Whole CanA Gene Amplification of Helicobacter pylori and Its Fingerprinting by Restriction Fragment Length Polymorphism

To set up a method of amplification for the whole CagA gene of Helicobacter pylori and its fingerprinting by restriction fragment length polymorphism(RFLP),nested PCR was employed in combination with TD-PCR to amplify the gene and EcoRI and Hind Ⅲ were used to generate the RFLP fingerprinting.Target DNA fragments from 13 of 20 samples were successfully amplified and the relevant RFLP fingerprintings were obtained.It is concluded that the method can be used to amplify the whole CagA gene and CagA gene has apparent diversity of RFLP profile.     

健康教育对宫颈糜烂妇女治疗依从性的研究

目的 探讨多种形式的健康教育对提高宫颈糜烂治疗依从性的作用。方法 将确诊为宫颈糜烂的120例患者随机分为研究组、对照组。均给予相应的治疗。研究组增加健康教育内容。两组患者同时观察复诊率、治愈率、对医疗服务的满意程度、疾病的认知情况。结果 研究组增加健康教育后，患者复诊率、治愈率、诊疗满意程度、疾病认知情况均较对照组高(P＜O．05)。结论 健康教育能提高宫颈糜烂患者的治疗依从性，获得较高的治愈率。     

微创手术治疗高血压脑出血

目的 探讨微创手术治疗高血压脑出血的临床疗效。方法 132例高血压脑出血分成微创手术组(68例)和传统开颅手术组(64例)，分析两组手术的特点和手术时机，比较两组手术治疗的疗效。结果 微创组术后GOS良好23例、中残24例、重残9例、植物生存3例、死亡9例；传统组术GOS良好16例、中残15例、重残12例、植物生存6例、死亡15例。两组超早期或早期手术均有良好的预后，而微创组效果更佳。结论 微创手术治疗高血压脑出血能明显提高临床疗效，降低病死率。     

后腹腔镜手术在泌尿外科的应用

目的　探讨后腹腔镜手术治疗泌尿系疾病的方法及疗效。方法　采用后腹腔镜手术治疗泌尿系疾病3 5例 ,其中肾上腺肿瘤及肾上腺囊肿切除 13例 ,肾囊肿去顶术 15例 ,精索静脉曲张高位结扎术 4例 ,UPJ成形术 1例 ,乳糜尿行肾蒂淋巴管结扎术 1例 ,肾切除 1例。结果　 3 5例手术全部成功。手术时间 3 0～ 13 0分钟 ,术中平均出血 2 0ml。术后住院时间 1.5～ 14天 ,平均 4.5天。结论　腹腔镜手术治疗泌尿系疾病 ,具有损伤小 ,住院时间短 ,恢复快 ,并发症少等优点 ,值得临床推广应用     

In vitro differentiation of human adipose-derived adult stromal cells into neuron-like cells in hippocampal astrocyte conditioned medium

BACKGROUND: At present, researches on differentiating from human adipose-derived adult stromal cells (hADASC) to neuron-like cells are focus on inducing by artificial-synthetic compound solution; however, hippocampal astrocyte conditioned medium (HCAM) can induce in vitro differentiation from hADASC to neuron-like cells is still unclear. OBJECTIVE: To observe whether HCAM can induce in vitro differentiation from hADASC to neuron-like cells. DESIGN: Randomized control study. SETTING: Department of Neurology, Taixing People's Hospital; Central Laboratory, North China Coal Medical College. MATERIALS: Donor of adipose tissue was donated by female volunteers suffering from caesarean section in the department of obstetrics & gynecology in our hospital and aged 20－35 years. Adipose tissue was collected from subcutaneous tissue of abdomen during the operation. In addition, 8 male newborn Wistar rats within 24 hours with average body mass of 20 g were provided by Animal Institute of Chinese Academy of Medical Sciences. Rabbit-anti-human Nestin polyclonal antibody, rabbit-anti-human glial fibriliary acidic protein (GFAP) polyclonal antibody, rabbit-anti-human neuro-specific enolase polyclonal antibody and mouse-anti-human microtubal associated protein 2 (MAP-2) polyclonal antibody were provided by Wuhan Boster Company. METHODS: The experiment was carried out in the Central Laboratory of North China Coal Medical College from October 2004 to June 2005. hADASC was cultured with HCAM and its growth and morphological changes were observed under inverted phase contrast microscope. Immunocytochemistry, immunofluorescence and Western blotting were used to evaluate the expressions of Nestin, which was a specific sign of nerve precursor, neuro-specific enolase and MAP-2, which was a specific sign of nerve cell, and GFAP, which was a specific sign of neuroglial cells. MAIN OUTCOME MEASURES: Nestin, which was a specific sign of nerve precursor, neuro-specific enolase and MAP-2, which was a specific sign of nerve cell, and GFAP, which was a specific sign of neuroglial cells. RESULTS: On the 3rd day of culture, partial hADASC started deformation from slender shuttle-shape cells to neuron-like cells. It suggested that cells stretched out apophysis, which were mainly double-pole or multiple-pole cells. Five days later, immunohistochemical detection suggested that expression of Nestin (10.5±0.037) was found out in cells; meanwhile, expressions of GFAP (38.4±0.052) and neuro-specific enolase (NSE) (15.7±0.023) were also found out in cells; however, expression of MAP-2 was not observed. Western blot indicated that, 5 days after effect of HCAM, Nestin was found out in hADASC; meanwhile, expressions of GFAP and neuro-specific enolase were also found out; however, expression of MAP-2 was not observed. CONCLUSION: HCAM can induce the differentiation from hADASC to neuron-like cells in vitro.     

Whole-brain 3D perfusion MRI at 3.0 T using CASL with a separate labeling coil.

A variety of continuous and pulsed arterial spin labeling (ASL) perfusion MRI techniques have been demonstrated in recent years. One of the reasons these methods are still not routinely used is the limited extent of the imaging region. Of the ASL methods proposed to date, continuous ASL (CASL) with a separate labeling coil is particularly attractive for whole-brain studies at high fields. This approach can provide an increased signal-to-noise ratio (SNR) in perfusion images because there are no magnetization transfer (MT) effects, and lessen concerns regarding RF power deposition at high field because it uses a local labeling coil. In this work, we demonstrate CASL whole-brain quantitative perfusion imaging at 3.0 T using a combination of strategies: 3D volume acquisition, background tissue signal suppression, and a separate labeling coil. The results show that this approach can be used to acquire perfusion images in all brain regions with good sensitivity. Further, it is shown that the method can be performed safely on humans without exceeding the current RF power deposition limits. The current method can be extended to higher fields, and further improved by the use of multiple receiver coils and parallel imaging techniques to reduce scan time or provide increased resolution.     

Effects of human amniotic fluid on cartilage regeneration from free perichondrial grafts in rabbits.

After the chondrogenic potential of free grafts of perichondrium was shown in several experimental studies, perichondrium has been used to reconstruct cartilage tissue in various clinical situations. This study investigates the effects of human amniotic fluid on neochondrogenesis from free perichondrial grafts in a rabbit model. Since this fluid contains high concentrations of hyaluronic acid, hyaluronic acid-stimulating activator, growth factors, and extracellular matrix precursors during the second trimester, it may have a stimulating effect on neochondrogenesis. Perichondrial grafts, measuring 20 x 20 mm2 were obtained from the ears of 144 New Zealand young rabbits and were sutured over the paravertebral muscles. The rabbits were randomly divided into three groups with 48 rabbits per group. In group 1, 0.3 ml human amniotic fluid, and in group 2, 0.3 ml saline were injected underneath the perichondrial grafts. Group 3 formed the control group in which no treatment was given. Histologically, neochondrogenesis was evaluated in terms of cellular form and graft thickness at 2, 4, 6, and 8 weeks after surgery. In group 1, the mature cartilage was generated quickly and the cartilage plate in this group was significantly thick and extensive when compared with groups 2 and 3 at 8 weeks ( p<0.05 ANOVA). In conclusion, our study shows that human amniotic fluid enhances neochondrogenesis from free perichondrial grafts. The rich content of hyaluronic acid and growth factors possibly participate in this result.