“津血同源”“肾为唾”的研究

以23例肾功能不全与20例肾功正常患者的唾液尿素氮、钠、钾、舌面pH值和血液的相应生化检查作对照。结果唾液尿素氮、钾与血尿素氮、钾成正相关性。证实了中医“津血同源”“肾为唾”的理论。     

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.     

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.     

Mass spectrometric analysis of the structure of gamma II bovine lens crystallin.

The amino acid sequence of bovine gamma II-crystallin has been verified by a combination of electrospray and fast atom bombardment mass spectrometry. The molecular weight of gamma II, isolated by gel filtration and ion exchange chromatography, was determined to be 20,967 +/- 3 by electrospray mass spectrometry. Another aliquot of gamma II was completely digested by trypsin in a medium of 20% CH3CN and 0.1 M Tris, pH 8.2. The tryptic peptides were separated by reversed phase HPLC and identified by their molecular weights, as determined by fast atom bombardment mass spectrometry (FABMS). The identification of each peptide was confirmed by digesting the peptide further to give new peptides whose molecular weights were also determined by FABMS and related to the proposed amino acid sequences. The data from both types of mass spectrometric analyses were consistent with the sequence previously proposed by Hay et al. (J. Biol. Chem. 1987, 146, 332-338), including threonine at position 119. The FAB mass spectrum of one HPLC fraction suggested that disulfide bonding between Cys 18 and Cys 22 was present in at least half the protein preparation. Whether the Cys 18/Cys 22 disulfide bond was present in native gamma II or was produced during isolation or enzymic digestion could not be determined from these studies. Samples that had been stored for several weeks showed that several of the cysteines had become disulfide bonded. These studies illustrate the power of mass spectrometric techniques to accurately confirm the primary structure of proteins and to identify post-translational modifications.     

兔动静脉短路环法诱导组织工程骨支架材料血管生成的初步研究

Objective To compare the effects of 2 vascular carriers, arteriovenous loop and arteri-ovenous bundle, on inducing angiogenesis in coral scaffold of vascularized tissue-engineered bone in animal models.Methods Thirty-six adult male New Zealand rabbits were randomized into 2 even groups.In group A, an arteriovenous loop (AVL) was formed by microsurgical anastomosis at the proximal ends between the femoral poptiteal artery and vein, and placed in the circular side groove of the coral block (6 mm × 8 mm × 10 mm) .In group B, flow-through vessels bundles of both femoral artery and vein were placed in the side grooves of the coral block.All the implants in 2 groups were wrapped by a micro-porous expand-ed-polytetrafluoroethylene (ePTFE) membrane, and fixed subcutaneously by suturing.Evaluation methods included gross morphological observations, histological examinations, India ink perfusion and vascular casting after 2, 4, 6 weeks.The density of blood vessels was analyzed by the statistical software SPSS 10.0.Results All the corals were encased by newly formed fibrovascular tissues in 2 groups.Ink-stained vessels distributed the surfaces and side grooves, and invaded the interspaces of corals.The degree of vascularization increased over the course of experiment.Blood vessel density demonstrated a significant continuous increase between 2 and 6 weeks after implantation in group A.The mean value of blood vessel density in group A (2 weeks 276.60±4.67, 4 weeks 517.20±10.66, 6 weeks 707.00 ±11.87) was significantly higher than in group B (2 weeks 153.60 ±7.16, 4 weeks 269.40±6.80, 6 weeks 279.20±6.53) (P <0.01).Vascular casting showed that in group A, significant blood vessels sprouted from all areas of the loop, espe-cially at the entrance of the arteriovenous pediele where the small tubes were densely interconnected.In group B, however, no blood vessels sprouted from the arteriovenous bundles and only some small vessels grew from the entrance and exit.Conclusions A vascularized coral model can be constructed by inserting an ar-teriovenous loop or an arteriovenous bundle, useful in vascular bone tissue engineering.The former, however, have stronger abilities to induce angiogenesis than the latter.     

Treatment of severe subglottic laryngotracheal stenosis using hyoid graft with sternohyoid muscle flap]

OBJECTIVE: To evaluate the possibility and reliability of the hyoid-sternohyoid graft transfer in the correction of server subglottic laryngotracheal stenosis, and delineate the operation skills and clinical results. METHODS: Seven patients with severe subglottic stenosis underwent laryngotracheal reconstruction using the hyoid grafts with sternohyoid muscle flaps (HG-SHMF). Five of these patients had traumatic subglottic stenosis, one with scar tissue of unknown etiology arising in the subglottic region, another with tracheal narrowing caused by inhalation of hydrochloric acid. RESULTS: All seven patients were successfully decannulated with moderate good voice. The average time from reconstruction to decannulation was 15.4 months. The stent was endoscopically removed with a range of 3 to 22 months; the mean time required for stenting was 9.6 months. Two patients who received additional salvage reconstruction procedures because of graft or stent displacement were extubated with improved voices and satisfactory airway. CONCLUSIONS: The HG-SHMF transfer was a single-stage reconstruction, relatively simple procedure that can restore an adequate airway and a good voice. Patients undergoing laryngotracheal reconstruction with HG-SHMF must have regular, long-term follow-up since graft displacement and recurrent granulation tissue or scar reformation can cause restenosis after an initially successful surgery. This procedure should be used in a large number of patients to further test its reliability.     

甘露醇和低温对兔骨骼肌缺血再灌流损伤的保护作用

通过兔后肢缺血模型，观察了甘露醇和低温对骨骼肌缺血再灌流损伤的保护作用。结果：甘露醇和低温可以不同程度地降低再灌流后患肢静脉回流血中细胞内酶（ＣＰＫ、ＬＤＨ、ＡＳＴ）的活性和脂质过氧化反应终末代谢产物丙二醛（ＭＤＡ）的含量，减轻骨骼肌细胞超微结构的破坏。两者单独应用效果不明显（Ｐ＞０．０５），联合应用效果非常明显（Ｐ＜０．０１）。提示自由基介导的脂质过氧化反应参与骨骼肌缺血再灌流损伤，甘露醇和低温对其具有协同保护作用。