Rapid detection of Mycobacterium tuberculosis in various clinical specimens by using polymerase chain reaction combined with a nonradioactive hybridization system

A DNA amplification system using the polymerase chain reaction (PCR) combined with a nonradioactive digoxigenin-labeled probe hybridization was employed to detect Mycobacterium tuberculosis in clinical specimens. One hundred and thirty specimens were tested by several methods including routine culture method, acid-fast staining, BACTEC 460 detection system, PCR, and PCR-hybridization techniques. Sixteen out of 130 specimens were culture positive on Middlebrook 7H11 agar, 10 were positive with acid-fast staining, 18 were positive with BACTEC 460 detection system, 23 were positive with PCR technique, and 62 were positive with PCR-nonradioactive hybridization technique. When compared with culture results, PCR-nonradioactive hybridization had an overall sensitivity of 100% (16/16) and a specificity of 59.7% (68/114). However, 28 out of 46 (60.9%) PCR-nonradioactive hybridization positive specimens which were culture negative had clinical data supporting the diagnosis of tuberculosis. In addition, 4 specimens which were negative by routine culture but positive by BACTEC 460 detection system and two specimens which were negative by routine culture but positive by acid-fast staining were all positive by PCR-hybridization technique. These data suggest that routine culture method may not be sensitive enough to detect M. tuberculosis in all kinds of clinical specimens. Taking this deviation into account, the specificity of PCR-nonradioactive hybridization technique may be rectified range from 63% (68/108) to 79.1% (68/86). PCR itself is not satisfactory enough to detect M. tuberculosis in specimens (the sensitivity and specificity were 56.3% and 87.7%, respectively) in this study. However, when it combines with DNA hybridization technique, they can be a very powerful and rapid diagnostic tool to detect M. tuberculosis in clinical specimens.     

Napsin A Expression in Subtypes of Thyroid Tumors: Comparison with Lung Adenocarcinomas

Napsin A is widely used in the diagnosis of lung adenocarcinoma and has also been reported to be positive in cases of thyroid carcinomas. We investigated napsin A levels through immunohistochemistry on whole sections of 210 primary thyroid tumors of various subtypes and another 41 metastatic thyroid carcinomas, and compared these with 125 primary and 25 metastatic lung adenocarcinomas. The results showed that napsin A was expressed in 23.8% thyroid tumors and 30.3% papillary thyroid carcinomas. Most cases showed a focal and weak to moderate expression. In comparison, 80.8% primary lung adenocarcinomas expressed napsin A, with mostly diffused and strong expression. For metastatic carcinomas of thyroid and lung origin, napsin A was detected in 39.0% of thyroid carcinomas in contrast to 88.0% in cases of lung adenocarcinomas. Comparisons of additional markers, TTF-1, CK7, thyroglobulin, and Pax-8 in metastatic carcinomas showed the overlapping expression of immunomarkers of TTF-1 and CK7. Thyroglobulin and Pax-8 were useful for distinguishing between metastatic carcinomas; however, Pax-8 may be a superior marker due to its higher sensitivity. The clinicopathological analysis of papillary thyroid carcinomas showed that the expression of napsin A was positively correlated with lymph node metastasis (p = 0.030). Here, we focused on the expression of napsin A in thyroid tumors and compared it with that in lung adenocarcinomas. The expression of napsin A is common in thyroid tumors and the combined expression of napsin A and TTF-1 in a metastatic thyroid carcinoma is a cause for concern due to chances of misdiagnosis as lung adenocarcinoma.

     

数字化和3D打印技术辅助治疗陈旧性髌骨骨折

目的探讨数字化和3D打印技术辅助陈旧性髌骨骨折精确截骨、复位和内固定方法,了解其在辅助治疗陈旧性髌骨骨折中的价值。方法自2006年以来,纳入5例陈旧性髌骨骨折畸形愈合患者采用CT薄层扫描、三维重建获得骨折部位骨关节三维模型,采用计算机辅助分析骨折移位和新生骨痂形成情况,CAD设计辅助手术模板引导骨折面准确截骨、精确复位,仿真模拟骨折复位、内固定过程,最后采用膝关节HSS评分标准评定术前、术后2周、末次随访时的膝关节功能。结果5例患者经14~23个月的随访,随访期间未发现螺钉、钢缆滑脱松动以及伤口感染等并发症,术后X线片证实骨折均解剖复位;根据膝关节HSS评分标准评价,术前HSS评分为47、53、35、40、42分,术后2周HSS评分为82、83、80、85、86分,末次随访HSS评分为95、86、89、90、91分。最后根据膝关节HSS评分标准评价:优5例。结论采用计算机辅助精确界定髌骨骨折面、模拟骨折复位内固定、个性化手术模板引导精确截骨、复位和内固定过程,解决了陈旧性髌骨骨折截骨部位确定困难、复位欠佳等问题。     

COVID-19疫情期急诊手足创伤患者诊治预案

新型冠状病毒肺炎(COVID-19)传染力强、人群普遍易感、潜伏期较长、部分患者早期表现隐匿的特点,本院制定相应的预警及诊治方案,为患者提供最优质的医疗服务诊疗过程中,最大限度地防止交叉感染、保护患者及家属身体健康。为了更好地配合保障疫情期间急诊手足创伤患者得到有效的救治,根据急诊手足创伤患者的特点和诊治原则,结合本院手足外科急诊的实际情况,制定COVID-19疫情期间急诊手足创伤患者诊治预案。     

A protective polymorphism in MMP16, improved blood gas levels,and chronic obstructive pulmonary diseases: Family and two population‐based studies

The aberrant expression of matrix metalloproteinases (MMPs) is known to contribute to the pathogenesis of airway remodeling and alveolar disruption in chronic obstructive pulmonary disease (COPD). In the discovery stage, 11 COPD from five families were subjected to whole‐genome sequencing, and 21 common polymorphisms in MMPs and TIMPs were identified. These polymorphisms were genotyped in two subsequent verification studies. Of these polymorphisms, c.2392G>A (rs2664370T>C) and c.4158C>A (rs2664369T>G) in MMP16 remained significantly different. Functionally, we found that MMP16 expression was significantly increased in peripheral blood monocytes (PBMCs) from COPD and in cigarette smoke extract‐treated 16HBE cells compared with controls. This was also shown by bioinformatics analysis. COPD carrying rs2664370CC showed decreased levels of MMP16 in the plasma and in PBMCs compared with those carrying CT and TT. Treatment with hsa‐miR‐576‐5p mimics led to a greater reduction in luciferase reporter activity in cells transfected with rs2664370CC. Moreover, blood levels of base excess, PCO₂, and PO₂ in COPD with rs2664370CC were significantly lower than those with rs2664370CT+TT. Taken together, these results demonstrate that the rs2664370T>C polymorphism in MMP16 protects against the risk of COPD, likely by favoring interaction with hsa‐miR‐576‐5p, leading to reduced MMP16 expression and improved blood gas levels.     

缺血性卒中患者脑运动区域协调性功能影像学研究

目的　评估缺血性脑卒中患者颅脑神经解剖学与脑区功能学的关联性,进一步挖掘缺血性卒中发生的神经机制。 方法　采集15例健康对照组和19例亚急性缺血性脑卒中上肢偏瘫患者,利用计算神经影像学计算个性化颅脑内部半球功能连接指标,采集治疗前后临床康复评分作统计相关分析。 结果　（1）与对照组相比,卒中患者大脑半球功能连接显著降低,差异有统计学意义(T=-10.077, P<0.001);（2）病灶异常集中于补充运动区（P=0.02）和中央前回(P=0.03),差异有统计学意义;（3）药物治疗前后患者组中央前回中的功能连接值与康复评分量表呈正相关,差异有统计学意义（r=0.29,P=0.03）。 结论　皮质下缺血性脑卒中患者大脑运动区域的半球间功能连接减少,可作为临床诊断和康复评估提供新的途径。     

时间相干电场的外周神经无损刺激

外周神经电刺激可用于运动康复和慢性神经痛治疗,但目前具有空间选择性的无损刺激仍是一个有待解决的问题。提出一种基于时间相干(TI)电场的外周神经选择性无损电刺激方法,对大鼠坐骨神经进行实验,在其大腿腹侧与背侧皮肤上以平行于神经的方向布置刺激电极,通过相干电场扫描,将TI刺激峰值定位到神经上进行选择性刺激。结果表明,该方法可以在预先不知道神经确切位置的情况下通过扫描得出将刺激电场作用到神经的最佳电参数,从而实现对神经的选择性无损刺激,而且在刺激作用点不变的前提下实现刺激强度的控制。在此基础上研究TI电场对大鼠坐骨神经的刺激阈值I_T,测量固定频差Δf=0.5 Hz(n=12),改变频率f=1～6 kHz与固定f=5 kHz(n=11),改变频差Δf=0.5～10 Hz下的I_T,并将其和等幅kHz电场(n=7)的I_T进行比较。结果表明,等幅kHz电场的I_T显著高于TI电场(P<0.05),而且不同频率f下的I_T也有显著性差异(P<0.05),而不同频差Δf下的I_T却没有显著性差异(P>0.05),说明TI电流对大鼠坐骨神经的I_T受f影响而不受Δf影响,且刺激阈值I_T与频率f成正比关系。     

A female with X‐linked Nephrogenic diabetes insipidus in a family with inherited central diabetes Insipidus: Case report and review of the literature

There are two forms of diabetes insipidus, central (neurohypophyseal), and nephrogenic, caused by pathogenic variants in the AVP gene and the AVPR2 or AQP2 genes, respectively. We report on a four‐generation family, seven individuals had central diabetes insipidus (CDI) and the female index patient seen from age 16 to 26 years had (mild) nephrogenic diabetes insipidus. In her father with CDI, a known pathogenic heterozygous AVP variant c.232_234del p.(Glu78del) was identified, confirming the diagnosis of CDI in him and the other affected family members. In the proband, molecular analysis disclosed a novel heterozygous AVPR2 gene variant, c.962A > T p.(Asn321Ile) and an extremely skewed X‐inactivation, confirming X‐linked nephrogenic diabetes insipidus (XL‐NDI). Whole exome sequencing showed no further causative mutation. This is the first report on the co‐existence of CDI and NDI in one family. Our review of symptomatic female AVPR2 heterozygotes includes 23 families with at least one affected female (including this study). There were 21 different causative mutations. Mutation types in females did not differ from those in males. Both severe XL‐NDI and mild forms were reported in females. All six females with severe XL‐NDI had complete loss‐of‐function (null) mutations. The remaining 17 female probands had milder XL‐NDI caused by 14 missense variants and three null variants of the AVPR2 gene. X‐inactivation was studied in nine of these females; all showed extreme or slight skewing. The review underlines that XL‐NDI in female AVPR2 heterozygotes is always accompanied by skewed X‐inactivation, emphasizing a need for X‐inactivation studies in these females.     

Frataxin is reduced in Friedreich ataxia patients and is associated with mitochondrial membranes

Friedreich ataxia is a progressive neurodegenerative disorder caused by loss of function mutations in the frataxin gene. In order to unravel frataxin function we developed monoclonal antibodies raised against different regions of the protein. These antibodies detect a processed 18 kDa protein in various human and mouse tissues and cell lines that is severely reduced in Friedreich ataxia patients. By immunocytofluorescence and immunocytoelectron microscopy we show that frataxin is located in mitochondria, associated with the mitochondrial membranes and crests. Analysis of cellular localization of various truncated forms of frataxin expressed in cultured cells and evidence of removal of an N-terminal epitope during protein maturation demonstrated that the mitochondrial targetting sequence is encoded by the first 20 amino acids. Given the shared clinical features between Friedreich ataxia, vitamin E deficiency and some mitochondriopathies, our data suggest that a reduction in frataxin results in oxidative damage.