Method for In Situ Evaluation of Superoxide Production by Pulmonary Macrophages in the Rat

In order to investigate superoxide production by pulmonary macrophages in the rat, a route was created by ligating both the inferior and superior venae cavae and resecting the aorta after cannulation through the inferior vena cava into the right atrium of the heart. Lung perfusion was performed via this route with nitro blue tetrazolium. Although there was no formazan deposition throughout the lung, it became detectable in both alveolar and interstitial macrophages when phorbol myristate acetate was added to the perfusate. This deposition was markedly enhanced by previous injection of Corynebacterium parvum. The deposition disappeared after further addition of Cu(Lys)₂, a scavenger of superoxide anions. This procedure may be useful for estimating in situ the ability of pulmonary macrophages to produce superoxide in the rat.     

Influence of Various Immunomodulators on the Induction of Experimental Autoimmune Encephalomyelitis in Lewis Rats

The effect of various immunomodulators on the induction of experimental autoimmune encephalomyelitis (EAE) is evaluated in the Lewis rat. Bordetella pertussis (BP) is the optimal inductor of EAE in this rat strain. Treatment of the animals with BP either before or after or simultaneously with guinea-pig spinal cord preparation (GpSC) resulted in an EAE about two weeks thereafter. Additional injection of living BCG, of CFA, IFA (incomplete Freund's adjuvant) or Vibrio cholerae neuraminidase (VCN) did not augment or mitigate the effect induced by BP or GpSC. Living BCG, IFA, VCN or Corynebacterium parvum (CP) did not induce EAE when given in combination with GpSC but without BP. CFA combined with GpSC only occasionally induced EAE. However, EAE could be induced by the combination of CFA and GpSC or IFA and GpSC in a part of the animals tested if they had been pretreated or simultaneously been injected with living BCG by intravenous route. EAE could not be enhanced by the additional injection of VCN. Surprisingly, most of the animals peracutely died after injection of CFA and BP in combination with GpSC when they had been pretreated with CP. This effect was most pronounced when pretreatment was done on day -4. No acute effect could be seen when CP was given simultaneously to CFA, BP and GpSC. Animals which did not peracutely succumb developed EAE similarly as those in the positive control groups. CP treatment simultaneously with BP but without CFA resulted in a reduction of the EAE specific mortality. This reduction could not be seen if treatment with CP was done after injection of GpSC and BP.     

The role of Ureaplasma parvum serovar-3 or serovar-14 infection in female patients with chronic micturition urethral pain and recurrent microscopic hematuria

BackgroundUreaplasma parvum (UP) is commonly isolated in the genitor-urinary tract and may cause various clinical features, including microscopic hematuria (MH). Some UP serovars are more commonly related with specific urogenital disease, but the evidences have been conflicting. This study primarily aimed to research the possible associations between specific UP serotypes and genito-urinary pathogenicity in female patients showing MH with/without chronic micturition urethral pain (CMP).MethodsThis study retrospectively reviewed 276 female patients having MH with/without CMP, who visited health screening center or female infertility clinic. All patients underwent multiplex polymerase chain reaction (PCR) tests with vaginal and urine samples to evaluate the infection rate and serotypes of UP. The antimicrobial susceptibility of UP and the predictors of CMP among UP infected patients were also analyzed. All patients were followed up at least for 6-months.ResultsForty-nine patients (17.8%) showed urinary UP infection. Urinary UP serotyping showed the prevalence of seorvar-1, -3, -6 and -14 were 24.5%, 30.6%, 18.4% and 26.5%, respectively. 79.6% of the urinary UP positive patients accompanied vaginal UP infection. 22 patients of the cohort (8.0%) had CMP whereas serovars-3 and -14 accompanied CMP in 54.5% and 41.0% cases, respectively. No serovars-6 infection case had CMP. 26.4% of the cohort were infertile whereas 10.9% of these infertile patients were positive for urinary tract infection with UP serotype-3 or -14. Doxycycline, josamycin and pristinamycin were the most active antibiotics with the lowest rate of resistance (0.0%) for treating UP. At 1-month post-initial treatment with doxycycline, all UP serotypes were eradicated and no patient complained of urethral discomfort. However, simultaneous urinary and vaginal reinfection of serovar-3 (5 cases) and serovar-5 (1 case) were confirmed at 3-months post-initial doxycycline therapy. The logistic regression analyses revealed that serovars-3 [hazard ratio (HR) 1.354, P value 0.018] and -14 (HR 1.103, P value 0.046) were significantly associated with CMP in female patients having MH.ConclusionsUP serovars-3 and -14 infections could be associated with CMP in female patients having MH. Doxycycline, josamycin and pristinamycin were effective for treating UP. Serovar-3 showed higher reinfection rate than other serotypes after antibiotics treatment.     

短棒状杆菌菌体细胞与细胞代谢产物生物学活性的比较

欲提高短棒状杆菌制剂的临床应用效果,为该产品进一步精制奠定基础,进行了短棒状杆菌有效成份的研究。在适宜的培养基中,以厌氧法培养短棒状杆菌,分别收获菌体及培养上清,做生物学活性检测。结果显示,菌体细胞内脾激活和抑瘤试验生物学活性显著,细胞代谢产物无生物学活性。实验证实,短棒状杆菌的有效成份在菌体细胞内,与细胞代谢产物无关。     

天津地区奶牛感染隐孢子虫情况的调查

目的调查天津市周边区县奶牛隐孢子虫感染情况。方法对5处奶牛粪样进行检查，采用改良抗酸染色法初筛，对检出的可疑阳性样本采用金胺-酚染色、金胺酚-改良抗酸染色、蔗糖不连续梯度离心法及饱和硫酸锌漂浮等方法进一步观察确定。结果个体养殖户奶牛粪检隐孢子虫阳性率西青区为8.33%（3/36），东丽区为70.00%（7/10），津南区为6.67%（1/15）。中型奶牛场奶牛粪样75份，其中武清县35份，蓟县40份，均为阴性。检出的隐孢子虫经初步鉴定分别为鼠隐孢子虫与微小隐孢子虫。结论天津地区奶牛有一定程度的隐孢子虫感染，需进一步加强小型养牛户的卫生防疫。     

非哺乳期肉芽肿性乳腺炎中亲脂性棒状杆菌的检测及抗菌药物的选择

目的探讨非哺乳期肉芽肿性乳腺炎患者乳腺穿刺液标本中亲脂性棒状杆菌的分布及对抗菌药物的敏感性。方法选择2017年9月至2018年5月该院收治的20例非哺乳期肉芽肿性乳腺炎患者为研究对象,收集经48 h培养阴性的乳腺穿刺液标本,通过延长培养时间及改变培养条件进行苛养菌的培养。采用16S rRNA测序分析和基质辅助激光解吸离子飞行质谱技术分别进行菌种的鉴定;采用E-test法对最低抑菌浓度(MIC)进行测定,采用欧洲药敏试验委员会(EUCAST)标准进行敏感性判读;采用1%吐温-80血平板进行对照的亲脂性试验。结果共分离得到20株苛养菌,其中16株克氏棒状杆菌,1株结核硬脂酸棒状杆菌,1株棒状杆菌某些种,2株厌氧菌(分别是黑尔嗜胨菌和贪婪丙酸杆菌)。1株结核硬脂酸棒状杆菌对所有试验药物均高度敏感,其余17株棒状杆菌体外对青霉素类、大多数头孢类药物敏感,但对红霉素、克林霉素高度耐药,少部分菌株对左氧氟沙星高度耐药。亲脂性试验显示,所有棒状杆菌均具有良好的亲脂性。结论亲脂性棒状杆菌,尤其是克氏棒状杆菌与非哺乳期肉芽肿性乳腺炎密切相关,亲脂性是其重要的致病因素。实验室须高度重视并改善培养条件及运用基质辅助激光解吸离子飞行质谱技术进行准确鉴定以提高培养检出率。临床治疗建议使用亲脂性较好且敏感的利奈唑胺等抗菌药物进行目标性治疗。     

微小隐孢子虫cDNA文库的构建及P23、CP15/60基因的克隆

目的　微小隐孢子虫地方株cDNA文库构建及P2 3、CP15 60基因的克隆。　方法　提取微小隐孢子虫总RNA、mRNA ,逆转录合成cDNA。将cDNA与 pUC18DNA连接 ,导入DH5α宿主细胞中生成cDNA文库。根据文献分别设计并合成两对PCR引物 ,从上述文库中筛选保护性基因 ,对PCR产物克隆、测序。　结果　文库容量为 1.9× 10 6个重组子 ,文库中cDNA插入片段大小介于 0 .4× 10 3～ 6.5× 10 3bp。从该文库中克隆出编码 2 3kDa、15 60kDa子孢子表面蛋白的核苷酸序列。　结论　成功地用 pUC18质粒载体构建了C .parvumcDNA文库。     

短小棒状杆菌制剂对小白鼠ICAM1/LFA1基因表达的研究

目的采用短小棒状杆菌制剂对小白鼠ICAM1/LFA1基因表达的研究,从而进一步研究短小棒状杆菌制剂对免疫调节和抗肿瘤机理深入研究提出依据。方法本试验采用realtime PCR方法,以GAPDH和-βactin分别作为各组织样品的内源控制基因。结果短小棒状杆菌制剂短小棒状杆菌制剂小白鼠腹腔注射之后,ICAM1基因在脾脏组织中上调,5 h出现高峰,短小棒状杆菌制剂小白鼠腹腔注射之后3-5 h LFA-1基因持续性上调。不同组织的LAF-1基因检测结果显示,小肠和脾脏均表现为高表达,尤以脾脏突出。结论短小棒状杆菌制剂的免疫机理和治疗肿瘤的作用与早期激动免疫细胞的第一信号有直接相关。     

Pentapeptide sharing between Corynebacterium diphtheria toxin and the human neural protein network

We describe the pentapeptides shared between the Corynebacterium diphtheria toxin and the human proteins associated with fundamental neural functions. We report that diphtheria toxin pentapeptides are spread among human antigens such as tuberous sclerosis proteins 1 and 2, reelin, contactin-4, neuroligins, semaphorin-5A, sodium channel protein type 1 subunit α, Williams–Beuren syndrome chromosomal region 1 protein, Williams–Beuren syndrome chromosomal region 20A protein. Williams–Beuren syndrome chromosomal region 8 protein, Bardet–Biedl syndrome 9 protein, Bardet–Biedl syndrome 10 protein, oligodendrocyte-myelin glycoprotein, neurofibromin-2, and periaxin. The data are discussed in relation to the bacterial immune escape phenomenon, and in the context of potential cross-reactions in diagnostic tests and immune therapies.     

The influence of nonspecific immunotherapy on the course of Murine Melanoma

BCG, Corynebacterium parvum, vitamin A, and levamisole were continuously administered to C57 BL/6 mice bearing B16 melanoma prior to, with, and following tumor implantation as well as before and coincident with surgical excision of the implanted tumor. Only the group given levamisole prophylactically for 8 weeks before tumor inplantation showed a significant difference from control mice but a tenfold increase in dosage in this drug did not exert a comparable effect.