High-level Expression and Purification of Human Zona Pellucida huZP3a^22-176 and huZP3b^177-348 Peptides in Escherichia coli

Objective To try making huZP3a^22-176 and huZP3b^177-348 polypeptides （representing an intact huZP^322-348 protein without its N-terminal signal peptide and C-terminal transmembrane domain ） express in E. coli at a higher level Methods The cDNAs encoding huZP3a and huZP3b were obtained with PCR method. The pBV221 plasmid was used to construct thermo-inducible recombinant expression vector. Purification of two target expression products employed an improved method of preparative gel polyacrylamide gel electrophoresis. Results Two polypeptides of recombinant huZP3a （rhuZP3a） and recombinant huZP3b （rhuZP3b） were all expressed respectively in an E. coli BL21（DE3）pLysS strain at a higher level, which were recognized by two specific polyclonal antisera in Western blotting test which recognize a linear B cell epitope present in rhuZP3a or rhuZP3b respectively. Using the shake-flask method, approximately 5 mg of rhuZP3a and rhuZP3b with more than 95% relative homogeneity were harvested from 1 L culture respectively. Conclusion The availability of two rhuZP3 polypeptides will help in detecting the immunogenicities of rhuZP3a and rhuZP3b through animal experiments and confirming the function domain of non-glycosylated huZP3 to induce acrosome reaction in vitro.     

区域性灾害医疗应急救援体系的建立与完善

借鉴“云娜”台风灾害中应急医疗救援实践的经验，论述了区域性灾害医疗应急救援体系建立的必要性及其建立与完善。为此，应建立医疗应急救援分级响应机制，提高急诊医疗服务体系在灾害事件中的整体应急反应能力，搞好区域性灾害医疗应急救援网络的战略布局，优化灾害医疗应急预案并有计划地组织演练，加强政府对灾害医疗应急救援的组织领导作用，统筹安排应急救援人力，提高救援人员自身的防范意识和能力。     

内蒙古自治区肾综合征出血热高发区实施疫苗接种防治策略的研究

内蒙古自治区(内蒙古)自1955年在呼伦贝尔市牙克石市北部的图里河镇发现首例肾综合征出血热(HFRS)病例以来，疫区逐年扩大。随着国产HFRS疫苗研制成功与推广应用，该疫苗安全性得到了现场证实；为此，我区制定用3年时间(2001—2003年)在HFRS高发区实施以疫苗接种为主的新防控对策，有效地降低了该病的发病率和死亡率。     

准分子原位角膜磨镶术后再近视的准分子激光上皮下角膜磨镶术治疗

目的探讨准分子激光上皮下角膜磨镶术(lasersubepithelial keratomileusis,LASEK)治疗准分子激光原位角膜磨镶术(excimer laser in situ keratomileusis,LASIK)后再近视的疗效及安全性。方法对21例(36眼)LASIK后再近视患者施行LASEK。LASEK术前的平均屈光度为(-1.98±0.65)D,平均角膜瓣下剩余基质厚度为(256±18.46)μm,LASEK平均切削深度为(36.58±11.24)μm。结果术后第6个月时裸眼视力为1.0以上者22眼(占61.1%),0.8以上者31眼(占86.1%),0.5以上者36眼(占100%),平均屈光度为(-0.18±0.53)D,未见最佳矫正视力下降,术后裸眼视力较术前明显提高,差异有显著性(P<0.05)。角膜瓣haze 0级者29眼(占80.6%),0.5级者5眼(占13.9%),1级者2眼(占5.6%)。结论LASEK治疗LASIK后再近视安全有效,但更长期的效果有待继续随访。     

壮尔颗粒对小鼠免疫功能的影响

目的观察壮尔颗粒对小鼠免疫功能的影响。方法采用氢化可的松造模法制作免疫抑制小鼠模型,进而观察壮尔颗粒对其腹腔巨噬细胞吞噬功能、溶血素形成、淋巴细胞转化的影响。结果壮尔颗粒能明显提高正常小鼠和免疫抑制小鼠巨噬细胞吞噬百分率、吞噬指数、血清溶血素含量和淋巴细胞转化率。结论壮尔颗粒对机体免疫功能有显著增强作用。     

二脱甲氧基姜黄素的降血脂和抗脂质过氧化作用

目的：探讨二脱甲氧基姜黄素（bisdemethoxy curcumin，Bdmc）降血脂、抗脂质过氧化作用和对脂代谢相关酶类活性的影响。方法：用高脂膳食喂饲Wistar大鼠，造成食饵性高脂血症后，用二脱甲氧基姜黄素和阳性对照药辛伐他汀进行实验性治疗。给药3周后处死动物，测定血清和肝脏总胆固醇（TC）、甘油三酯（TG）和血清高密度脂蛋白（HDL-C）含量，计算低密度脂蛋白（LDL-C）含量，同时测定肝素化血浆脂解酶（post-heparin lipolytic activity，PHLA）、脂蛋白脂酶（LPL）和肝脂酶（HL）的活性及血清丙二醛（MDA）含量．结果：二脱甲氧基姜黄素和辛伐他汀均能使血清和肝脏TC、TG含量降低，血清HDL-C含量升高、LDL-C含量降低、二脱甲氧基姜黄素能显著提高PHLA活性，对提高LPL和HIL的活性作用也较明显；二脱甲氧基姜黄素能有效降低MDA的含量，结论：二脱甲氧基姜黄素具有降低肝脏和血清脂质，特别是有显著的降TG和提高TG代谢相关酶活性的作用。     

V806M突变型Axin基因真核表达载体的构建及其在神经胶质细胞瘤内的表达

目的构建V806M突变型Axin基因真核表达载体pIRES2-EGFP-MT-Axin(V806M),并稳定表达于大鼠神经胶质瘤细胞系C6。方法用分子克隆技术,构建真核表达载体pIRES2-EGFP-MT-Axin(V806M),经NheⅠ和SmaⅠ双酶切鉴定后,用脂质体法稳定转染神经胶质瘤细胞C6。结果构建了真核表达载体pIRES2-EGFP-MT-Axin(V806M),稳定转染后,经荧光显微镜和免疫细胞化学染色法检测,可见细胞内有EGFP及Axin的表达。结论成功构建真核表达载体pIRES2-EGFP-MT-Axin(V806M),并在神经胶质瘤细胞C6中表达,为研究此种突变体Axin是否影响细胞的生物学行为以及是否参与胶质瘤的发生发展奠定了实验基础。     

胸段食管癌三维适形放疗摆位误差研究

Objective To study the setup errors in three-dimensional conformal radiotherapy (3DCRT) for thoracic esophageal carcinoma using electronic portal imaging device(EPID) and calculate the margins from CTV to PTV. Methods Forty-one patients with thoracic esophageal carcinoma who received 3DCRT were continuously enrolled into this study. The anterior and lateral electronic portal images (EPI) were aquired by EPID once a week. The setup errors were obtained through comparing the difference between EPI and digitally reconstructed radiographs(DRR). Then the setup margins from CTV to PTV were calculat-ed. By using self paired design,22 patients received definitive radiotherapy with different margins. Group A: the margins were 10 mm in all the three axes;Group B: the margins were aquired in this study. The differ-ence were compared by Paired t-test or Wilcoxon signed-rank test. Results The margins from CTV to PTV in x,y and z axes were 8.72 mm, 10.50 mm and 5.62 mm, respectively. Between the group A and group B, the difference of the maximum dose of the spinal cord was significant(4638.7 cGy±1449.6 cGy vs. 4310.2 cGy±1528.7 cGy; t=5.48, P=0.000), and the difference of NTCP for the spinal cord was also significant (4.82%±5.99% vs. 3.64%±4.70%;Z=-2.70,P=0.007). Conclusions For patients with tho-racic esophageal carcinoma who receive 3DCRT in author's department,the margins from CTV to PTV in x, y and z axes were 8.72 mm, 10.50 mm and 5.62 mm, respectively. The spinal cord could be better protected by using these setup margins than using 10 mm in each axis.     

成人脂肪间充质干细胞体外定向诱导分化为心肌样细胞的实验观察

目的:探索成人脂肪组织来源的间充质干细胞(ADMSCs)体外分离、培养方法,并化学诱导使其向心肌细胞分化,为心肌再生医学提供理想的种子细胞来源. 方法:体外分离成人ADMSCs并进行传代培养,5-氮胞苷(5-aza)诱导其向心肌细胞分化. 分别于诱导后14,21,28 d时用免疫细胞化学染色、RT-PCR法进行心肌细胞鉴定. 结果:ADMSCs经5-aza诱导后14 d时细胞形态趋向于心肌细胞,诱导28 d免疫细胞化学显示心肌特异性肌钙蛋白I(cTN-I)、结蛋白(Desmin)表达阳性,RT-PCR检测心肌β-肌球蛋白重链(Cardiac β-MHC)基因表达阳性. 结论:成人脂肪组织存在丰富的间充质干细胞且易于分离扩增,体外5-aza诱导可使其向心肌细胞分化,可做为心肌再生医学的理想种子细胞.     

体重指数与高血压患者脂肪肝的关系分析

目的针对高血压患者观察脂肪肝与体重指数的关系。方法对202例高血压患者行B超检查及实验室生化检测，根据超声影像学诊断分为脂肪肝组（98例）和非脂肪肝组（104例），同时检查（年龄、身高、体重、收缩压、舒张压）和实验室检查，将两组间一般资料进行t检验、卡方检验及多因素相关分析，观察高血压合并脂肪肝组和非合并脂肪肝组体重指数（BMI）的差异，及在不同体重指数人群中高血压患者脂肪肝的检出率和脂肪肝发生的危险因素。结果（1）高血压合并脂肪肝组与非合并脂肪肝组体重指数差异有显著性。（2）随着体重指数的增高，高血压患者脂肪肝的检出率呈增高趋势。（3）Logistic回归分析表明，在高血压患病人群中，扣除其他影响因素，BMI、性别（SEX）、丙氨酸氨基转氨酶（SLAT）是脂肪肝的独立危险因素，其中体重指数比数比（OR值）最高，而肥胖组的OR值较超重组OR值成倍增长。结论体重指数可能和高血压患者的脂肪肝发生有相关性。