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.     

槲皮素的结构修饰及生物活性研究进展

槲皮素是蔬菜、水果、中药中常见的黄酮类化合物,是天然的抗氧化剂,具有抗癌、抗糖尿病、抗菌、抗炎、抗病毒等生物活性。由于槲皮素生物利用度低,限制了其在临床上的应用。通过各种方法设计和合成新的槲皮素衍生物,以改善其缺点,进而发挥预防和治疗疾病的作用。通过对槲皮素衍生物的合成及其抗癌、抗糖尿病、抗炎、抗菌和抗病毒活性进行综述,并对其构效关系进行分析,为天然化合物的开发和利用奠定基础。     

安脑片联合奥拉西坦治疗脑卒中后血管性痴呆热毒内盛证临床研究

目的：观察安脑片联合奥拉西坦治疗脑卒中后血管性痴呆（VD）热毒内盛证的临床疗效。方法：按随机数字表法将100例脑卒中后VD热毒内盛证患者分为治疗组与对照组,每组50例。对照组采用奥拉西坦治疗,治疗组给予安脑片联合奥拉西坦治疗,2组均治疗4周。比较2组临床疗效与不良反应发生率。采用美国国立卫生研究院卒中量表（NIHSS）、简易智能精神状态检查量表（MMSE）、临床痴呆评定量表（CDR）、日常生活活动能力量表（ADL）评估神经功能、认知功能、痴呆程度和日常生活活动能力;比较双侧大脑中动脉（MCA）、基底动脉（BA）、大脑前动脉（ACA）、椎动脉（VA）平均血流速度,以及血清脑源性神经营养因子（BDNF）、血管内皮生长因子（VEGF）水平。结果：治疗后,治疗组临床疗效总有效率96.00%,高于对照组82.00%(P<0.05)。治疗后,2组NIHSS、CDR评分均较治疗前降低（P<0.05）,MMSE、ADL评分均较治疗前升高（P<0.05）;治疗组NIHSS、CDR评分均低于对照组（P<0.05）,MMSE、ADL评分均高于对照组（P<0.05）。治疗后,2组...     

Clinicopathological characteristics,prognosis, and chemosensitivity in patients with metastatic upper tract urothelial carcinoma

PurposeTo investigate the clinical characteristics, chemosensitivity, and outcome of metastatic upper tract urothelial carcinoma (UTUC).Patients and MethodsRecords of patients with metastatic UTUC since January 2005 were retrieved from a database that included clinical and survival data. Statistical analyses including survival and multivariate analyses of factors were respectively performed by the Kaplan-Meier method and Cox proportional hazard model.ResultsA total of 250 consecutive UTUC cases were evaluated. There were 56 patients (22.4%) with initially diagnosed stage IV disease. The most common metastatic sites were lung (39.6%), distant lymph nodes (39.2%), bone (19.6%), liver (18.0%), and adrenal gland (7.2%), respectively, and the local recurrence rate was 10.4%. Two hundred thirteen patients received first-line chemotherapy. The overall response rate was only 28.7% and the median progression-free survival time was only 5.0 months. The overall survival time of the cohort was 18.0 months. Multivariate analyses showed that initially diagnosed stage IV disease, number of metastatic organs ≥3, no response to chemotherapy and cycles of chemotherapy ≤2 were adverse prognosticators for overall survival.ConclusionUTUC presented to be more prone to metastasize than locally recur and thought to have low chemosensitivity. Stage IV disease at initial diagnosis, number of metastatic organs, response and cycles of chemotherapy were independent prognosticators for metastatic UTUC.     

安宫牛黄丸治疗基底节区脑出血患者颅内高压疗效观察及对血清B 型钠尿肽水平的影响

目的：观察安宫牛黄丸治疗基底节区脑出血患者颅内高压疗效及对氧化应激指标以及血清B 型钠 尿肽（BNP） 水平的影响。方法：选取80 例痰热内闭证基底结节区脑出血患者,按随机数字表法分为观察组 及对照组各40 例,对照组接受脑出血颅内高压常规方案治疗,观察组在对照组基础上联合安宫牛黄丸治疗。 比较2 组治疗前后颅内压（ICP）、平均动脉压（MAP）、心率（HR）、中医证候积分、总抗氧化能力（T-Aoc）、超氧 化物歧化酶（SOD） 以及BNP 水平的变化。结果：治疗后,2 组ICP、MAP 水平均较治疗前下降（P＜0.05）, HR 水平均较治疗前上升（P＜0.05）;观察组ICP、MAP 水平均低于对照组（P＜0.05）,HR 水平高于对照 组（P＜0.05）。治疗后,剧烈头痛、动作迟缓、嗜睡中医证候积分均较治疗前下降（P＜0.05）,观察组上述 3 项中医证候积分均低于对照组（P＜0.05）。治疗后,2 组T-Aoc、SOD 水平均较治疗前上升（P＜0.05）,BNP 水平均较治疗前下降（P＜0.05）;观察组T-Aoc、SOD 水平均高于对照组（P＜0.05）,BNP 水平低于对照 组（P＜0.05）。结论：安宫牛黄丸治疗痰热内闭证基底节区脑出血患者,能有效改善颅内高压,缓解氧化应激 反应,促进心脏功能的恢复。     

不同机制大动脉粥样硬化型脑卒中基线特征和血管内治疗后结局比较

目的 本研究旨在比较颅内大血管伴/不伴同侧颈内动脉闭塞的大动脉粥样硬化型脑卒中患者基线特征以及行血管内治疗后结局的差异。方法 对DIRECT-MT亚组进行回顾性分析,以比较前循环大动脉粥样硬化（large-artery atherosclerosis,LAA）型卒中串联闭塞和颅内闭塞接受血管内治疗（endovascular treatment,EVT）的患者的基线特征和预后,分析不同机制学特征（动脉粥样硬化或动脉-动脉栓塞）对临床结局的影响。结果 LAA型卒中患者共108例,其中串联闭塞63例,颅内闭塞45例。颅内闭塞组患者高血压史率高于串联闭塞组（77.8% vs. 52.4%, P=0.007）。颅内闭塞组闭塞部位最常见于大脑中动脉M1段（88.6%）,而串联闭塞组颅内闭塞主要位于颈内动脉颅内段（49.2%）和大脑中动脉M1段（49.2%）（P＜0.001）。两组患者在年龄、性别、术前抗栓、他汀类药物的使用,卒中、房颤、吸烟史,基线mRS、NIHSS评分,是否静脉溶栓,侧枝循环,以及救治流程时间差异均无统计学意义（P均＞0.05）。90天mRS 0-2分的患者比例两组差异无统计学意义（53.3% vs. 41.9%, P=0.243）。颅内闭塞组术后成功再灌注率高于串联闭塞组（93.3% vs. 77.4%, P=0.026）,但术后24-72小时血管再通的比例前组低于后组（57.1% vs. 77.2%, P=0.034）。最终梗死体积,颅内闭塞组小于串联闭塞组（20.1 vs. 34.5, P=0.025）。术后NIHSS评分,90天EQ-5D-5L评分和BI指数等其他次要结局,两组间差异无统计学意义（P均＞0.05）。两组在90天内的死亡率,发生的无症状性和症状性颅内出血率,5-7天时在另外的血管区域新发脑梗死,以及新流域栓塞的患者百分比相似,差异无统计学意义（P均＞0.05）。结论 动脉粥样硬化导致的串联闭塞相较于孤立颅内闭塞,末次造影成功再灌注率较低,梗死体积更大,但术后24-72小时再通率更高,且神经功能良好预后率以及不良事件发生率均与颅内闭塞相仿。