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1.
目的 基于核酸试纸条技术,建立一种快速检测蜡样芽胞杆菌entFM毒素基因的方法。 方法 煮沸法提取蜡样芽胞杆菌DNA,以蜡样芽胞杆菌entFM毒素基因作为靶基因,采用NCBI primer-BLAST 5.0软件设计特异性引物并通过克隆转化鉴定PCR产物,建立并组装核酸试纸条,评价核酸试纸条的特异性、灵敏度和稳定性。 结果 蜡样芽胞杆菌DNA浓度为300 mg?L-1,纯度约为1.60。PCR产物阳性条带经切胶回收、克隆转化和测序比对,与GenBank数据库中已登记的entFM相似性为100%。在pH值7.0条件下,每100 μL 胶体金溶液加入3.3 μg链霉亲和素浓度标记,质控线浓度为1.8 g?L-1,检测线浓度为1 g?L-1时,硝酸纤维素膜上质控线与检测线均可与PCR产物反应出现清晰的红色条带。按照最适条件组装成核酸试纸条,PCR产物6 μL,样品展开液100 μL,检测10 min后可观察结果。核酸试纸条特异性与电泳结果一致,仅蜡样芽胞杆菌为阳性结果,与金黄色葡萄球菌、铜绿假单胞杆菌、大肠埃希菌和沙门氏菌皆无交叉反应,为阴性结果;灵敏度检测,核酸试纸条DNA质量浓度同时降至10-3 mg?L-1仍可准确检测,普通PCR电泳结果显示DNA质量浓度同时降至10-1 mg?L-1出现目的条带,核酸试纸条比普通PCR灵敏度提高100倍;稳定性检测,核酸试纸条在第6、9和12个月进行检测稳定性一致。 结论 建立的核酸试纸条检测蜡样芽胞杆菌entFM毒素基因灵敏度高、特异性强且具有良好稳定性,适用于快速鉴别蜡样芽胞杆菌entFM毒素基因。  相似文献   
2.
《Vaccine》2022,40(30):4038-4045
PurposeAs protection from COVID-19 following two doses of the BNT162b2 vaccine showed a time dependent waning, a third (booster) dose was administrated. This study aims to compare the antibody response following the third dose versus the second and to evaluate post-booster seroconversion.MethodsA prospective observational study conducted in Maccabi Healthcare Services. Serial SARS-CoV-2 Spike IgG tests, 1,2,3 and 6 months following the second vaccine dose and one month following the third were obtained. Neutralizing antibody levels were measured in a subset of participants. Per individual SARS-CoV-2 Spike IgG titer ratios were calculated one month after the booster administration compared to titers one month following the second dose and prior to booster.ResultsAmong 110 participants, 56 (51%) were women. Mean age was 61.7 ± 1.9 years and 66 (60%) were immunocompromised. One month after third dose, IgG titers were induced 7.83 (95 %CI 5.25–11.67) folds and 2.40 (95 %CI 1.90–3.03) folds compared to one month after the second, in the immunocompromised and immunocompetent groups, respectively. Of the 17 immunocompromised participants who were seronegative after the second dose, 4 (24%) became seropositive following the third. Comparing the titers prior to the third dose, an increase of 50.7 (95 %CI 32.5–79.1) fold in the immunocompromised group and 25.7 (95 %CI 19.1–34.7) fold in and immunocompetent group, was observed.ConclusionA third BNT162b2 vaccine elicited robust humoral response, superior to the response observed following the second, among immunocompetent and immunocompromised individuals.  相似文献   
3.
目的 预测“十四五”期间我国城乡人口和基层医疗卫生机构“2 + X”家庭医生团队数量,探究我国城乡家庭医生服务覆盖情况,为家庭医生健康发展提供科学依据。方法 依据2013—2019年相关数据,采用GM(1,1)灰色预测模型预测2020—2025年我国城乡人口数和“2 + X”家庭医生团队数量和相关卫技人员数。观察每 2000 服务人口标准下,2018—2025年我国城乡“2 + X”家庭医生团队服务人数和服务覆盖率。结果 2020—2025年我国城镇人口数逐年增长,到2025年增至98 831万人;城市基层医疗卫生机构全科医生数、注册护士数、家庭医生团队数量逐年增长,到2025年分别增至171 712人、848 324人、171 712个;全科医生与护士比逐年上升,到2025年升至1∶4.94;2018—2025年我国城市“2 + X”家庭医生团队服务人口覆盖率逐年上升,到2025年升至35%。2020—2025年我国乡村人口数逐年下降,到2025年降至48 053万人;乡镇基层医疗卫生机构全科医生数、注册护士数、家庭医生团队数量逐年增长,到2025年分别增至447 672人、854 976人、447 672个;全科医生与护士比逐年下降,到2025年降至1:1.91;2018—2025年我国乡村“2 + X”家庭医生团队服务人口覆盖率逐年上升,理论上2022年实现全面覆盖。结论 2018—2025年我国城乡“2 + X”家庭医生团队数量逐年增加,但城市家庭医生缺口较大,应加强全科医学人才培养,增强家庭医生职业吸引力,缩小城乡全科医生薪资差距,优化家庭医生服务覆盖均衡性。  相似文献   
4.
IntroductionAlthough hyperferritinemia may reflect the inflammatory status of patients with non-alcoholic fatty liver disease (NAFLD), approximately 33% of hyperferritinemia cases reflect real hepatic iron overload.AimTo evaluate a non-invasive method for assessing mild iron overload in patients with NAFLD using 3T magnetic resonance imaging (MRI) relaxometry, serum hepcidin, and the expression of ferritin subunits.MethodsThis cross-sectional study assessed patients with biopsy-proven NAFLD. MRI relaxometry was performed using a 3T scanner in all patients, and the results were compared with iron content determined by liver biopsy. Ferritin, hepcidin, and ferritin subunits were assessed and classified according to ferritin levels and to siderosis identified by liver biopsy.ResultsA total of 67 patients with NAFLD were included in the study. MRI revealed mild iron overload in all patients (sensitivity, 73.5%; specificity, 70%). For mild (grade 1) siderosis, the transverse relaxation rate (R2*) threshold was 58.9 s?1 and the mean value was 72.5 s?1 (SD, 33.9), while for grades 2/3 it was 88.2 s?1 (SD, 31.9) (p < 0.001). The hepcidin threshold for siderosis was > 30.2 ng/mL (sensitivity, 87%; specificity, 82%). Ferritin H and ferritin L subunits were expressed similarly in patients with NAFLD, regardless of siderosis. There were no significant differences in laboratory test results between the groups, including glucose parameters and liver function tests.ConclusionsMRI relaxometry and serum hepcidin accurately assessed mild iron overload in patients with dysmetabolic iron overload syndrome.  相似文献   
5.
Human auditory ossicles, the malleus, the incus, and the stapes, are located in the tympanic cavity in the temporal bone and through forming a chain for the sound transmission from the tympanic membrane to the cochlea, they play an important role in the hearing process. Despite their clinical, phylogenetic, and evolutionary significance, the morphometry of the human ear bones has not been examined systematically. The ear ossicles are the smallest bones of the human skeleton, attaining their final size and morphology already at birth. Initially, they have been found to exhibit minimal morphometric variation, but further studies brought the opposite results. The aim of this study was to examine the morphometric variation of human auditory ossicles recovered from medieval and postmedieval subadult skeletons from Poland, Central Europe. The analysis involved in a total of 166 ear bones. Their measurements were performed on microscopic images using CorelDraw x4, according to a protocol of Quam and Rak with modification of Flohr et al. and Wadhwa et al. Our study showed a significant metric variation in the measurements taken at areas of the greatest morphological variability of the ossicles. We found that greater linear dimensions were associated with lower values of angular measurements. These results reveal the inherent variation found in these supposed functionally constrained structures. Representation of even greater number of populations, time periods, and developmental stages are needed. Further study will expand our understanding of the global scope of variation found in ear ossicular morphology and its functional implications for paleoanthropology.  相似文献   
6.
ObjectivesOur aim was to evaluate the effect of the updated European Organization for Research and Treatment of Cancer (EORTC) and Mycoses Study Group 2019 definitions for invasive pulmonary aspergillosis (IPA) on patient classification and the related all-cause 12-week mortality.MethodsIn this retrospective cohort study from our tertiary care centre, we reclassified patients with haematological malignancy who underwent bronchoalveolar lavage between 2014 and 2019 for suspected IPA using the novel EORTC 2019 criteria. We performed receiver operating characteristic curve analysis to define the optimal cut-off for positive PCR and galactomannan and present survival analyses and their possible association with these diagnostic criteria through post hoc comparisons with log rank and Cox regression.ResultsFrom 323 episodes of suspected IPA in 282 patients, 73 were reclassified: 31 (42.5%) from possible to probable IPA, 5 (6.8%) from EORTC criteria not met to probable IPA, and 37 (50.7%) from EORTC criteria not met to possible IPA. Probable IPA increased therefore 11.1% (64/323, 19.8% to 100/323, 30.9%), mostly due to positive PCR (31/36, 86.1%). There was no difference in mortality between newly defined possible and probable IPA (log rank p = 0.950). Mortality was higher in probable cases with lower cycle thresholds (Ct values) versus higher Ct values (p = 0.004). Receiver operating characteristic curve analysis showed an optimal Ct value cut-off of 36.8 with a sensitivity of 75% (95% CI 64.9%–85.1%) and a specificity of 61.7% (95% CI 53.5–69.9) for 12-week mortality.DiscussionThe new EORTC criteria led to 11.1% more probable IPA diagnoses, mostly due to Aspergillus PCR. Restricting positive PCR to below a certain threshold might improve the discrimination of the new EORTC IPA categories for mortality.  相似文献   
7.
目的建立一种基于规律成簇的间隔短回文重复序列及其相关蛋白(CRISPR/Cas13a)的乙型肝炎病毒(HBV)共价闭合环状DNA(HBV cccDNA)检测方法。方法提取2017年6月至2020年10月于首都医科大学附属北京佑安医院就诊的4例乙型肝炎患者肝脏总DNA后,使用HindⅢ内切酶和质粒安全性ATP依赖DNA酶(PSAD)分别进行酶切;根据松弛环状DNA(rcDNA)和cccDNA的结构差异,设计特异性扩增HBV cccDNA的引物,对酶切后的产物进行滚环扩增(RCA)和PCR扩增;并筛选crRNA,建立基于CRISPR/Cas13a技术的HBV cccDNA检测新方法。结果利用α-1-抗胰蛋白酶(A1AT)和HBV表面抗原(HBsAg)引物对双重酶切后的产物进行扩增,验证产物中HBV基因组的存在;利用HBV cccDNA和HBV rcDNA引物对PSDA酶切后的产物扩增,验证了产物中只存在HBV cccDNA;利用RCA后的阳性样本作为模板梯度稀释,然后进行PCR扩增转录后使用CRISPR/Cas13a检测,计算出检测下限为10拷贝/μl。结论本研究建立了RCA-PCR-CRISPR-Cas13a的新型检测方法,可对HBV cccDNA进行高灵敏度和高特异性检测,为乙型肝炎患者抗病毒治疗评估、治疗终点的确定以及调整治疗方案提供了有效的监测手段。  相似文献   
8.
9.
目的 通过解剖学观察,揭示颈椎间孔内口区锚链韧带的形态及其分布特征,探讨其与神经根型颈椎病神经卡压之间的关系。 方法 12具成人脊柱颈段防腐标本,正中矢状切开,从脊神经根袖根部切断根袖,在外科显微镜下解剖观察C3/4~C7/T1椎间孔内口处神经根袖周围的锚链韧带,记录韧带的形态、分布、起止点及走行。 结果 120个椎间孔内口脊神经根袖周围共发现560条锚链韧带,所有韧带均呈放射状连于神经根袖与椎间孔内口周围骨膜壁,两端附着紧密,不易分离。各椎间孔内口韧带的数目均≥4个。锚链韧带形态主要包括带形和索形两种。带形韧带258条,宽度(4.5±2.6)mm(4.1~5.2 mm),索形韧带302条,直径(2.5±1.8)mm(1.2~3.8 mm)。C3/4、C7/T1椎间孔内口区韧带较为松散纤细,数量较少;C4/5~C6/7椎间孔内韧带较为坚韧而粗壮,数量较多。 结论 颈椎椎间孔内口区神经根周围存在锚链韧带,将神经根锚定于周围椎间孔骨膜壁,极大限制了神经根自由移动范围,可能与突出椎间盘一起导致神经根卡压,是神经根型颈椎病的潜在解剖学因素。  相似文献   
10.
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