Comparison of amplified Q beta replicase and PCR assays for detection of Mycobacterium tuberculosis.

Because of the long time required to isolate Mycobacterium tuberculosis in culture, there is an acute need for simple rapid methods for direct detection of M. tuberculosis from human sputum specimens. We have developed and characterized quantitative manual Q beta replicase and PCR assays for M. tuberculosis. The Q beta replicase assay was based on reversible target capture of M. tuberculosis 23S rRNA followed by amplification of a replicatable detector probe with Q beta replicase. For PCR assays, primers generating a 370-bp amplification product from the IS6110 insertion element were used in combination with a control plasmid containing an internal deletion in the IS6110 amplicon. Serial dilutions of M. tuberculosis were spiked into sputum and subjected to digestion and decontamination with N-acetyl-L-cysteine and NaOH. Assay conditions were optimized for hybridization and sample processing chemistries in order to maximize sample utilization. Following assay optimization, the sensitivities of the Q beta replicase and PCR assays of spiked sputum samples were 0.5 and 5.0 CFU per assay reaction, respectively. The effects of sputum matrix on each assay were examined by testing 20 patient sputum samples which had been cultured for M. tuberculosis. The culture-positive samples included smear-positive and smear-negative samples. The results of the Q beta replicase assay were not inhibited by sputum and were in 100% agreement with those of culture, including detection of 10 culture-positive specimens. However, using an internal control plasmid coamplified with each PCR as an indicator, we detected PCR inhibition in 9 of 20 samples tested.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of a neurotoxigenic Clostridium butyricum strain isolated from the food implicated in an outbreak of food-borne type E botulism.

Neurotoxigenic Clostridium butyricum was isolated from the food implicated in an outbreak of clinically diagnosed type E botulism in China. PCR assay showed that the isolate (LCL 155) contained the type E botulinum toxin gene. This appears to be the first report of neurotoxigenic C. butyricum causing food-borne botulism.     

Pharmacological basis for use of madecassoside in gouty arthritis: anti-inflammatory,anti-hyperuricemic,and NLRP3 inhibition

Objectives: Gouty arthritis is caused by the deposition of monosodium urate (MSU) crystals in joints, which is associated with the rise of serum urate content. This study aims to investigate the therapeutic effect of Madecassoside on gouty arthritis and hyperuricemia.

Methods: DBA/1 mice were intradermally injected with MSU to stimulate joint inflammation or intraperitoneally injected with MSU to trigger peritonitis. Moreover, ICR mice were exposed to potassium oxonate to stimulate hyperuricemia.

Results: Madecassoside repressed MSU-triggered pad swelling, joint 99mTc uptake, and joint inflammation in DBA/1 mice with gouty arthritis. Neutrophil infiltration and IL-1β & IL-6 & MCP-1 secretion was also alleviated in lavage fluids from DBA/1 mice with peritonitis due to Madecassoside treatment. Furthermore, Madecassoside decreased MSU-induced neutrophil cytosolic factor 1, caspase-1 and NLRP3 expression in mice with peritoneal inflammation. In hyperuricemic mice, Madecassoside improved renal dysfunction. Serum uric acid, BUN, and creatinine were down-regulated by Madecassoside.

Conclusion: These findings indicate that Madecassoside has potential to ameliorate inflammation in both acute gouty arthritis model and peritonitis model, probably via regulating IL-1β and NLRP3 expression.

Practical point: Madecassoside also exhibited a urate-lowering effect and a renal protective effect in hyperuricemic mice.     

The outer membrane of Pasteurella multocida 3:A protects rabbits against homologous challenge.

The protective efficacy of a vaccine purified from the Pasteurella multocida 3:A outer membrane (OM) was evaluated in rabbits by homologous challenge. Twenty-seven rabbits were divided into four groups: 1, vaccinated with OM and challenged; 2, nonvaccinated and challenged; 3, vaccinated with OM only; and 4, nonvaccinated and not challenged. Rabbits were immunized intranasally with 1 mg of OM protein on days 0, 7, 14, and 35, challenged intranasally on day 49, and killed on day 63. Mortality rates were 0, 67, 0, and 0% for groups 1 through 4, respectively. The prevalence of pneumonia was reduced from 73 (group 2) to 20% (group 1). The severity of pneumonia was reduced from 0.62 (group 2) to 0.07 (group 1), as measured by the group lesion index. The number of P. multocida in nasal cavities was reduced from 3.89 x 10(5) (group 2) to 6.19 x 10(2) (group 1). The geometric mean number of P. multocida in lungs was 8,360,000-fold less in group 1 than in group 2. Similarly, the prevalence of P. multocida colonization in nonrespiratory organs was reduced from 47 (group 2) to 4% (group 1). Furthermore, group 1 and 3 rabbits developed significantly elevated immunoglobulin A antibodies in nasal secretions and lung lavages and significantly elevated immunoglobulin G antibodies in lung lavages and sera. In addition, rabbit immune sera contained antibodies against P. multocida OM proteins and lipopolysaccharides and inhibited P. multocida proliferation in mouse lungs. These results indicate that a vaccine prepared from the OM of P. multocida provides a significant protection in rabbits against homologous challenge.     

Molecular modification of anticholinergics as probes for muscarinic receptors. 2. Amino esters of alpha-methyltropic acid

As a continuation of our goals to study molecular probes for muscarinic cholinergic receptors, a series of 3-substituted 2-methyl-2-phenylpropanoates with the general structure of C6H5C(CH2X)(CH3)COOCH2CH2NEt2 where X = OH, OTs, F, Cl, Br, I, and OAc were prepared and their antispasmodic activities examined on isolated rat ileum preparations. Structure-activity relationship studies with these compounds provide further evidence suggesting that binding of an aromatic moiety in a specific location within the hydrophobic region of the receptor is important for anticholinergic potency. A nucleophilic displacement of chloride by "naked" fluoride under mild conditions is also reported.     

Radiographic findings in cotton textile workers and the relationship to cigarette smoking

A group of 140 cotton textile workers from Shanghai, Shandong, Guangxi, and Beijing have had their chest radiographs taken using similar requirements. Most had come from the preparatory departments of cotton mills and had a history of exposure to cotton dust for at least 20 years. As controls, 140 healthy individuals with no dust exposure were matched with respect to sex, age, and smoking history. All the radiographs were read according to the ILO International Pneumoconiosis Classification, and the manifestations belonging to categories 0/0 and 0/1 were grouped as "normal," whereas categories 1/0, 1/1, and 1/2 and above as "abnormal." There was no significant difference in X-ray abnormalities between cotton textile workers and controls (P greater than 0.05). But when the data on the cotton textile workers and controls were combined, an appreciable difference in the incidence of radiographic abnormalities was found between smokers and non-smokers (P less than 0.001). Cotton dust exposure may induce nonspecific interstitial pulmonary changes, but these changes were exaggerated by cigarette smoking. These two factors appeared to have an additive effect on the pulmonary X-ray findings.     

ALPHA-FETOPROTEIN VARIANTS AND THEIR CLINICAL SIGNIFICANCE

The molecular heterogeneity of serum alpha-fe- toprotein (AFP) in 36 patients was studied by lectin affinity-radioimmunoelectrophoresis. Two variants of AFP, concanavalin-A (Con-A) reactive and the Con-A nonreactive, were obtained. AFP in all 13 patients with hepatoma consisted of the two va- riants, but the Con-A reactive variant was the ma jor one. In 8 patients with benign liver disorder, however, only the Con-A reactive variant of AFP was detected. Over 50"70 0f AFP in 5 patients with yolk sac tumor was Con-A nonreactive. Thus, there were three different patterns of AFP variants. The clinical implications are discussed.     

遗尿儿童和青少年情绪和行为问题及相关因素研究

目的 调查河南省儿童青少年遗尿症的情绪和行为问题的现况及其影响因素。 方法 从2019年10月—2020年3月,采用分层随机整群抽样方法抽取河南某县5所中小学校,采用一般资料调查表、长处和困难问卷及儿童睡眠调查表进行调查,对影响遗尿患儿情绪和行为的因素进行单因素、多因素分析。 结果 实际发放问卷4 500份,回收4 120份,有效问卷3 912份,有效回收率为94.95%。调查结果显示有7.82%儿童患有遗尿症。遗尿组的情绪症状、多动和困难总分得分明显高于正常组(P＜0.001),两组品行问题、同伴交往和社会行为得分的比较差异无统计学意义(P＞0.05)。Pearson相关分析结果显示,遗尿儿童和青少年的困难总分与打鼾、夜间呼吸问题、嗜睡、注意力缺陷、睡眠总分均存在正相关关系(P＜0.05)。多元线性回归结果显示睡眠呼吸紊乱、照顾者文化程度低、便秘、学习成绩较差是遗尿患儿情绪和行为问题的影响因素。 结论 遗尿患儿情绪和行为问题的检出率高,影响因素是睡眠呼吸紊乱、照顾者文化程度低、孩子学习成绩差、便秘,应引起关注。     

手术室护理质量控制指标构建与应用

目的构建手术室护理质量控制指标,为持续改进手术室护理质量提供依据。方法基于文献回顾、小组讨论、专家函询、全国护理质量促进联盟审议等确定手术室护理质量控制指标。于2021年10月-12月在全国19家三甲医院手术室进行试应用。结果手术室护理质量控制指标涵盖结构指标2项、过程指标5项、结果指标1项。19家医院手术室护士与手术间比例中位数为3.2:1,工作日手术间平均开放时间为9.89 h,手术安全核查正确执行率、手术物品清点正确执行率、外科手消毒正确率、手术标本管理规范合格率、手术间环境卫生消毒合格率、术中2期及以上压力性损伤发生率的合计率分别为83%、86%、83%、88%、95%、0%。结论手术室护理质量控制指标为评价手术室护理质量提供了工具,为持续改进手术室护理质量提供了参考。     

儿童体质发展情况分析

目的 对昆明市城区连续三年3~6岁学龄前儿童体质测试结果进行分析,寻找学龄前儿童体质发展中的不平衡点,为制定体格锻炼、全面提高学龄前儿童身体素质的相关措施提供建议。 方法 选取2017-2019年连续进行体质测定的昆明市城区幼儿园38所,采用SPSS 20.0统计软件对结果进行数据分析。相关指标包括:身高、体重、10米折返跑、立定跳远、网球掷远、双脚连续跳、坐位体前屈、走平衡木8个体质项目,以及年龄别身高、年龄别体重、身高别体重3个生长发育评价结果。 结果 2019年体质测定的优秀率从2018年的32.5%提高到35.7%(χ²=62.512,P<0.05)。坐位体前屈0分值比率从2018年的1.7%上升到2019年的3%;除10米折返跑外,其余五个项目5分值比率2019年较2018年上升了1.8%~8.8%。与正常儿童6个项目相比,身高低常儿童平均分低1.855分(t=-16.416,P<0.05),体重低常儿童平均分低1.856分(t=-17.346,P<0.05);体重超常儿童平均分低0.277分(t=-2.798,P<0.05),消瘦儿童平均分低0.917分(t=-6.073,P<0.05),肥胖儿童平均分低0.571分(t=-5.490,P<0.05);身高超常儿童平均分高0.812分(t=7.123,P<0.05),且这生长发育异常的儿童在6个体质项目表现中,大部分项目的低分(0~2分)比率高于正常儿童。 结论 重视儿童柔韧性、平衡性锻炼,加强对家长的健康教育及认识的提高,做好儿童健康管理,做到早期预防及治疗生长发育相关疾病,才能持续促进体质健康发展,保障儿童健康。