提取青蒿素实验条件的研究

对从青蒿叶中提取青蒿素的各种方法及实验条件进行实验比较，结果表明，采用索氏提取法及以Ｂ号油为溶剂，具有提取率较高，时间短、操作方便、耗油少等特点。     

骨形成蛋白(BMP)复合材料的实验研究

作者利用异种、同种、半脱钙骨基质明胶和羟基磷灰石四种材料植入20只大鼠肌肉内,通过光学显微镜观察比较了各种材料的吸收速度、抗原性和成骨情况。结果表明,异种骨基质明胶在植骨早期有轻度的排斥反应。羟基磷灰石不易被吸收,无诱导成骨作用。同种骨基质明胶适合作为较小骨缺损的骨形成蛋白复合材料,修复较大骨缺损,半脱钙骨基质明胶是较佳的复合材料。     

“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。     

对IgM肾病的探讨

50例肾活检的原发性肾小球肾炎中,发现6例IgM肾病。主要病理特点为系膜增殖及IgM荧光沉积。临床表现以肾病综合征为特点,血清IgM升高,蛋白尿及血尿,并呈高凝状态。     

中药泡洗治疗糖尿病足90例疗效分析

目的为了观察中药泡洗对糖尿病足的疗效。方法对90例糖尿病足患者用中药（方剂组成：桃仁、红花、桂枝、没药、乳香、花椒、川芎等各10g）水煎制后分为2袋各300ml配合足浴仪泡洗，2次，d，30min／次，10d为1疗程。结果全组90例患者显效66例，有效24例，总有效率100％。结论中药泡洗治疗糖尿病足疗效确切。     

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.     

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.