冷冻切片常遇到的问题及解决对策

目的 提高制作冷冻切片质量，确保实验结果科学性。方法 通过对切片机工作室温度设置选择，防卷板的调整应用及切片常遇到的问题分析，阐述提高制作高质量冷冻切片的方法。结果 通过对冷冻切片机有关参数的设定，提高了切片质量。结论 冷冻切片常遇到的问题，通过及时调整切片机相应设置，可以便捷解决。     

Expression of canine interferon-gamma by a recombinant vaccinia virus and its antiviral effect

A recombinant vaccinia virus-expressing canine interferon (IFN)-gamma (vv/cIFN-gamma) was constructed. In rabbit kidney (RK13) and canine A72 cells infected with vv/cIFN-gamma, IFN activity was detected in the culture supernatants of both cell types. Canine IFN-gamma was also detected in both cell extracts by Western blot. The activity of the recombinant canine IFN-gamma in RK13 cells was higher than that in A72 cells. The vv/cIFN-gamma could not grow in A72 cells at a low multiplicity of infection, probably due to the antiviral activity of the canine IFN-gamma produced. Although exogenous IFN-gamma did not inhibit the growth of vaccinia virus, addition of anti-canine IFN-gamma serum recovered the growth of the vv/cIFN-gamma on A72 cells in a dose-dependent manner. These results suggest that the growth of vv/cIFN-gamma was inhibited by IFN-gamma produced in a paracrine and autocrine manner. In addition, the recombinant canine IFN-gamma inhibited the multiplication of canine herpesvirus, pseudorabies virus and canine adenovirus type 1 in Madin-Darby canine kidney cells. The antiviral effect of canine IFN-gamma was more effective than that of canine IFN-beta. From the present studies, we concluded the recombinant virus may be a useful suicide viral vector.     

Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay

To isolate Babesia equi genes encoding immunodominant proteins, a cDNA expression library prepared from B. equi mRNA was immunoscreened with B. equi-infected horse serum. Eighteen positive cDNA clones were obtained, and the clone that showed the strongest immunoreactivity, designated Be82, was further characterized. The Be82 gene consisted of 1,953 bp and contained a partial open reading frame lacking the 5'-terminal sequence. As shown by Western blot analyses, immune sera from mice intraperitoneally injected with the Be82 gene product recognized the 82- and 52-kDa proteins of B. equi but not those of Babesia caballi. The glutathione S-transferase fusion protein expressed in Escherichia coli that was purified and used as the antigen in the enzyme-linked immunosorbent assay reacted specifically with B. equi-infected horse sera. These results suggest that the Be82 gene product is a potential diagnostic antigen candidate in the detection of B. equi infection in horses that will be useful both in the performance of epidemiological studies and in the granting of quarantine passes.     

Construction of Canine Herpesvirus Vector Expressing Foreign Genes Using a lacZ-TK Gene Cassette as a Double Selectional Marker

An improved method for constructing canine herpesvirus (CHV) recombinants expressing foreign genes by using the lacZ-TK gene cassette as a double selectional marker was developed. A recombinant CHV carrying the lacZ-TK gene at a targeted gene locus was constructed and used as a parental virus for generating new recombinants. The parental virus formed blue plaques and was sensitive to TK-specific drugs, while newly generated recombinants, in which the lacZ-TK gene was replaced with the desired foreign gene, become both resistant to the TK-specific drugs and formed white plaques. Recombinants were isolated by using the combination of drug selection and color selection. This improved method allows construction of recombinant CHV with great ease, because the drug selection can enrich the frequency of recombinant CHV from 0.01–0.1% to 10–80%. This method was employed to construct a recombinant CHV that expressed rabies virus (RV) glycoprotein (G protein). This revised version was published online in July 2006 with corrections to the Cover Date.     

采用MDA-MB-231细胞膜进行表皮生长因子受体活性的检测

目的　探讨能否直接利用肿瘤细胞膜进行表皮生长因子受体 (Epidermalgrowthfactorreceptor,EGFR)催化活性检测。方法　首先筛选出EGFR基因表达水平相对较高的细胞株MDA MB 2 31,通过差速离心制备细胞膜 ,采用Westernblotting检测EGFR催化磷酸化的程度。结果　底物被磷酸化 ,加入特异性拮抗剂AG14 78后 ,磷酸化被抑制。结论　利用肿瘤细胞膜检测EGFR活性的设想是成立的 ,并且其方法简便、经济。     

肝移植病人的ICU期观察及护理

目的：通过对8例肝移植、1例肝肾联合移植术后护理经验的总结,介绍肝移植病人的护理重点及方法;方法：隔离室准备、神经系统及生命体征监测、排斥反应监测、预防感染及心理护理;结果：肝移植术后早期（ICU）的护理至关重要,直接关系到近远期疗效和生存率;结论：新技术的开展要求高素质的护理人员和高质量的护理水平。     

两个地区东方田鼠基因组RAPD分析比较研究

为了建立一种与临床上冠脉搭桥手术和经皮腔内冠状动脉成形术（PTCA）等病理过程相似的血管再狭窄模型,探讨血管再构和新内膜形成的机理,及在血管再狭窄中所起的重要作用,我们用不同品系（BALB／cA和C57BL／6J）小鼠的左颈总动脉进行了结扎,在2周和4周后,分别作病理学观察,结果发现两种品系小鼠产生了不同的病理结果,C57BL／6J小鼠有明显的血管狭窄和炎症细胞浸润,而BALB／cA小鼠虽有炎症细胞的浸润,但血管的内膜无炎症细胞浸润,也无血管狭窄表现,结果提示：血管再狭窄是血管再构和新内膜形成的共同原因。     

介入联合手术治疗膀胱癌32例疗效分析

目的观察髂内动脉灌注化疗联合手术治疗膀胱癌的疗效。方法对32例中晚期膀胱癌先行介入治疗,再手术切除。结果32例膀胱癌完全缓解(CR)率为71．9％。结论介入治疗中晚期膀胱的手术提供了良好基础,对一部分原本行扩大切除术甚至全切的患实行单纯部分切除术,从而保留了膀胱生理功能,有助于提高患生活质量,延长生命。