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中华生物医学工程杂志
中华生物医学工程杂志

钟南山

双月刊

1674-1927

cjbme@vip.tom.com

020-81340157

510182

广东省广州市东风西路195号广州医学院内28栋1楼

中华生物医学工程杂志/Journal Chinese Journal of Biomedical EngineeringCSTPCD
查看更多>>中华医学会、广州医学院主办。本刊办刊宗旨：密切关注并报道生物医学工程学研究的新理论、新方法、新技术，跟踪生物医学工程学在临床中的最新应用成果，服务广大临床医生，促进生物医学工程学的学科发展。生物医学工程的发展一直是临床医学进步的动力，而临床医学所需要解决的问题则是生物医学工程创新的源泉。临床医生、科学家和产业界工程技术人员的紧密合作将为人类创造更美好的健康长寿的新生活。内容和栏目：（1）编者导读；（2）专论；（3）论著；（4）新技术与临床；（5）新技术研发；（6）生物技术；（7）综述；（8）专题讲座。
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    狂犬病毒糖蛋白修饰外泌体构建神经靶向磁共振分子探针的实验研究

    张帆刘晨鹭严彩英童明敏...
    1-7页
    查看更多>>摘要：目的 利用狂犬病毒糖蛋白（RVG）修饰的外泌体与超顺磁性氧化铁纳米颗粒（SPION）构建神经靶向分子探针RVG-Exo-SPION，研究其表征及体外磁共振成像（MRI）效果。 方法 构建RVG-Lamp2b质粒并转染HEK293细胞，提取膜表达RVG的外泌体，电穿孔与SPION合成磁性分子探针RVG-Exo-SPION。采用免疫印迹、透射电镜分析（TEM）、纳米粒径追踪分析（NTA）及体外毒性分析（CCK-8法）方法分别对分子探针外泌体膜蛋白表达、形状、尺寸和生物相容性进行分析和鉴定，并通过激光共聚焦成像、普鲁士蓝染色和MRI联合评估Neuro-2A细胞对分子探针的摄取能力。 结果 双酶切法证明目的基因重组至pcDNA3.1质粒中，免疫印迹显示重组质粒被有效转染至HEK293细胞，外泌体标记蛋白CD9和CD63呈高表达。TEM显示修饰和电穿孔后的外泌体保持其正常的形状、尺寸和膜结构，外泌体电穿孔后能见到SPION的存在。NTA显示工程化的外泌体粒径分布窄，粒径分布峰值为140 nm。Neuro-2A细胞与1~100 mg/L不同浓度梯度的RVG-Exo-SPION共同孵育时，细胞的活力差异无统计学意义（均P>0.05）。激光共聚焦显微成像分析显示外泌体能高效地进入细胞，线形图显示实验组荧光强度更高；普鲁士蓝染色验证了RVG-Exo-SPION对Neuro-2A细胞的靶向能力，靶向组的阳性细胞标记率明显高于对照组[（75.8±5.6）%比（19.1±2.5）%），P<0.05]；体外MRI显示灰阶值在不同铁浓度（5、10、25、50、100 mg/L）之间有明显区别，且在Fe≥25 mg/L的浓度下易于识别。 结论 基于RVG修饰外泌体构建的磁性分子探针RVG-Exo-SPION在体外实验中具有较好的稳定性和神经靶向性，为进一步用于体内研究提供了可行性。 Objective To construct a nerve-targeting molecular probe, RVG-Exo-SPION, by binding rabies virus glycoprotein (RVG) modified exosome to superparamagnetic iron oxide nanoparticles (SPION), characterize the probe, and validate its performance on magnetic resonance imaging (MRI) in vitro. Methods The RVG-Lamp2b plasmid was constructed and transfected into HEK293 cells. Exosomes expressing RVG on their membrane were extracted, electroporated, and bound with SPION to generate the magnetic molecular probe RVG-Exo-SPION. Western blotting, transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and in vitro cytotoxicity assay (CCK-8 method) were used to analyze or verify the expression, shape, size and biocompatibility of proteins on exosomal membrane of the molecular probe. The uptake of molecular probe by Neuro-2A cells was evaluated by confocal laser scanning microscopy (CLSM), Prussian blue staining and MRI. Results Double enzyme digestion confirmed that the target gene was cloned into the pcDNA3. 1 plasmid. Western blotting showed that the recombinant plasmid was effectively transfected into the HEK293 cells as reflected by high expression of exosomal marker proteins CD9 and CD63. TEM showed that the modified and electroporated exosomes maintained their normal shape, size and membrane structure. Presence of SPION could be seen after electroporation of the exosomes. NTA showed that the engineered exosomes had a narrow distribution range of particle size with a peak at 140 nm. Co-incubated with RVG-Exo-SPION at concentrations from 1 to 100 mg/L did not incur significant difference in viability of Neuro-2A cells (all P>0.05). CLSM showed efficient influx of exosomes into the Neuro-2A cells, with higher fluorescence intensity indicated in line graph for the RVG-Exo group compared with the controls. Prussian blue staining verified the Neuro-2A-targeting ability of RVG-Exo-SPION which yielded higher rate of positive cell labeling compared with the control group[ (75. 8±5. 6) % vs (19. 1±2. 5) %,P< 0.05]. In vitro MRI showed significant differences in gray scale values across varying iron concentrations (5, 10, 25, 50, and 100 mg/L), which could be readily distinguished at iron concentrations of 25 mg/L and above. Conclusion The magnetic molecular probe RVG-Exo-SPION constructed with RVG-modified exosome shows good stability and nerve-targeting ability in vitro, and this underpins the rationale for further studies in vivo.

    外泌体狂犬病病毒糖蛋白氧化铁磁性纳米颗粒磁共振成像药物靶向分子探针

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    松质骨的衰减和弥散——超声引导脊柱导航的意义

    李索远李培洋邵维维焦阳...
    8-16页
    查看更多>>摘要：目的 结合Micro-CT扫描数据了解椎体松质骨的声学特性，以此作为脊柱融合手术中超声导航后路椎弓根螺钉固定的理论依据。 方法 以2块牛脊椎松质骨块和2块人同种异体脊椎松质骨块模拟椎弓根螺钉通道内松质骨的情况，用Micro-CT扫描获得各骨标本的CT参数，用中心频率分别为2.2、2.5、3、12 MHz的无聚焦宽频换能器，分别对其进行超声透射实验，通过水听器检测穿过骨块样品的超声波振幅、声衰减和声速数据，计算骨块样品对不同频率超声波的衰减及声速影响。 结果 高密度人同种异体骨的骨体积分数和骨小梁数高于低密度人同种异体骨（均P<0.05），骨小梁分离度和结构模型指数低于低密度人同种异体骨（均P<0.05）。4块骨样本在4个不同频率下，超声波振幅随骨块厚度的增加而减小（均P<0.05），声衰减随骨块厚度的增加而增大（均P<0.05）。随着频率的增加，低频超声（2.2、2.5、3 MHz）下2 mm薄牛松质骨的声衰减逐渐减小[（r（牛骨1）=0.95，r（牛骨2）=0.89，均P<0.05]，4块骨样本声速均逐渐增大[r（牛骨1）=0.71，r（牛骨2）=0.81，r（高密度人同种异体骨）=0.35，r（低密度人同种异体骨）=0.61，均P<0.05]。牛松质骨中，在相同的低频超声（2.2、2.5、3 MHz）下，声速随骨厚度的增加而增加（均P<0.05）。 结论 松质骨骨块的CT参数特征和多种频率超声透射下的声学特性具有相关性，为超声导航系统的研发奠定了理论基础。 Objective To understand the acoustic profiles of vertebral cancellous bone in ultrasonography using data of Micro-CT scanning as the reference, so as to lay a theoretical foundation for ultrasound-guided posterior pedicle screw fixation in spinal fusion surgery. Methods Two cancellous bovine spinal bone blocks and two human cancellous allogeneic spinal bone blocks were used to simulate the cancellous bones along the pedicle screw channel. Each bone specimen was obtained of CT parameters on Micro-CT scanning, then subjected to ultrasound transmission measurement using an unfocused broadband transducer with central frequencies of 2.2 MHz, 2.5 MHz, 3 MHz, and 12 MHz, respectively. The acoustic amplitude, attenuation, and velocity of the ultrasound penetrating each bone specimen were captured by a hydrophone. Then the attenuation and impact on acoustic velocity of ultrasound by the bone specimens under different ultrasonic frequencies were calculated. Results High-density human allogeneic bone blocks presented higher bone volume fraction (BV/TV) and greater trabecular number (TB. N) (both P<0.05), lower trabecular separation and structural model index (bothP<0.05), compared with the low-density human allogeneic ones. For each of the 4 bone specimens under the 4 different frequencies, the ultrasound amplitudes decreased (allP<0.05) and the acoustic attenuation in creased (allP<0.05) with greater bone thickness. With rising frequency, a downward reduction in acoustic attenuation of low-frequency ultrasound (2.2 MHz, 2.5 MHz and 3 MHz) was noted inside the 2 mm thin cancellous bovine bone (r=0.95 for bovine bone block #1, r=0.89 for bovine bone block #2, both P<0.05), and an upward increase in acoustic velocity was noted inside all of the 4 bone blocks (r=0.71 for bovine bone block #1, r=0.81 for bovine bone block #2, r=0.35 for the high-density human allogeneic bone block, r=0.61 for the low density human allogeneic bone block, all P<0.05). In cancellous bovine bone blocks tested with any given low frequency of the ultrasound (2.2 MHz, 2.5 MHz, and 3 MHz), the acoustic velocity increased with greater bone thickness (allP<0.05) . Conclusion There is a correlation between CT parameter features of cancellous bone blocks and their acoustic profiles with ultrasound transmission at varying frequencies. This finding theoretically implies a foundation for development of ultrasound navigation systems.

    超声检查外科手术,计算机辅助脊柱松质骨

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    基于传输的超声造影序列度量学习用于甲状腺结节的识别

    刘春蕊万鹏孔文韬张道强...
    17-25页
    查看更多>>摘要：目的 建立甲状腺超声造影血流灌注相似性度量学习方法，对临床常见的甲状腺结节进行分类诊断。 方法 收集2017年1月至2020年12月南京鼓楼医院145例甲状腺结节超声造影病例，经手术病理明确诊断，包括20例结节性甲状腺肿，33例甲状腺滤泡状腺瘤，68例甲状腺微小乳头状癌和24例甲状腺乳头状癌。引入时序约束最优传输构建甲状腺超声造影度量学习（TCSML）方法，自动对齐局部造影增强模式并量化为动态血流灌注差异，并与已有的12种度量学习或多视图学习方法比对并辅助甲状腺结节进行分型鉴别。 结果 在与已有的12种度量学习或多视图学习方法的对比中，该TCSML模型的平均诊断准确度75.14%，灵敏度68.85%，精准度78.85%和F1分数69.79%，高于其他学习方法（均P<0.05）。模型的最优输入序列长度为7，最佳时序正则系数为0.05，最优特征投影维度为40，最优结构化范数约束为0.004。 结论 该模型能够有效辅助临床医生初步判断甲状腺分型类别，并提供主要的局部增强模式，为超声医生提供直观地诊断参考。 Objective To establish a metric learning approach that measures the similarity in blood perfusion on contrast-enhanced ultrasound (CEUS) of the thyroid gland, and to validate its use in categorical diagnosis of common thyroid nodules in clinical settings. Methods A total of 145 patients with thyroid nodules who underwent CEUS in Nanjing Drum Tower Hospital between January 2017 and December 2020 were included. According to post-surgical pathology, these patients comprised 20 cases of nodular goiter, 33 of follicular thyroid adenoma, 68 of papillary thyroid microcarcinoma, and 24 of papillary thyroid carcinoma. A temporally regularized optimal transport algorithm was introduced to develop an approach of thyroid contrast-enhanced sonography metric learning (TCSML) that automatically aligned local patterns of contrast enhancement and quantified the dynamic difference in blood perfusion. Then we compared TCSML to other 12 available approaches of metric learning or multi-view learning for the diagnostic performance in classification of thyroid nodules. Results Compared with the other 12 available approaches of metric learning or multi-view learning, TCSML on average yielded a diagnostic accuracy of 75.14%, a sensitivity of 68.85%, an accuracy of 78.85% and an F1 score of 69.79%, outperforming its counterparts (all P<0.05). The TCSML model presented an optimal input sequence length of 7, optimal temporal regularization coefficient of 0. 05, an optimal feature projection dimension of 40, and an optimal structured norm constraint of 0.004. Conclusion The TCSML model can effectively help clinicians in preliminary classification of thyroid nodules and, with a highlight on major modes for local enhancement, provides an intuitive diagnostic reference for sonographists.

    甲状腺结节超声心动描记术,多普勒,彩色血管造影术度量学习方法最优传输时序对齐虚拟序列

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    烟酰胺核糖对脂毒性心肌病的影响及机制探讨

    谢赛阳方文熙刘士强邓伟...
    26-33页
    查看更多>>摘要：目的 探讨烟酰胺核糖对肥胖相关脂毒性心肌病的影响及其具体机制。 方法 将C57BL/6N雄性小鼠（8周龄）分为标准饮食组（对照组）、高脂饮食组和高脂+烟酰胺核糖饮食组（n=10），高脂饮食组通过高脂饲料喂养24周构建肥胖小鼠脂毒性心肌病模型，高脂+烟酰胺核糖饮食组小鼠在高脂饲料的基础上在饮用水中加入40 mmol/L烟酰胺核糖喂养12周。造模结束后检测小鼠尾动脉血压，收集小鼠外周血分析血糖、血脂等指标，应用超声心动图评估小鼠心功能，随后进行小鼠心脏取材并用于病理学和分子生物学分析。分别通过心脏大体拍照检测小鼠心重与胫骨长、麦胚凝集素（WGA）定量小鼠心肌细胞面积和苦味酸-天狼猩红（PSR）染色显示小鼠心肌纤维化水平评估烟酰胺核糖对小鼠心肌重构的影响。采用透射电镜检测小鼠心肌脂滴浸润情况；采用免疫印迹检测小鼠心肌细胞Sirt1及Serca2a蛋白表达水平。 结果 与对照组比较，高脂饮食小鼠的体质量、收缩压、舒张压、总胆固醇、三酰甘油和LDL-c水平均明显增高（均P<0.05），应用烟酰胺核糖后可缓解高脂所致的上述指标的增高（均P<0.05）；高脂饮食和应用烟酰胺核糖均不影响小鼠心率和血糖。与对照组比较，高脂饮食组小鼠左心室射血分数和心脏超声E峰、A峰比值（E/A）降低（均P<0.05），烟酰胺核糖则增高左心室射血分数和E/A比值（均P<0.05）。与对照组比较，高脂饮食组小鼠心重与胫骨长比值、左心室心肌细胞横截面积、胶原含量均增加（均P<0.05），烟酰胺核糖则可抑制小鼠心肌的上述肥大和纤维化表现（均P<0.05）。与对照组比较，高脂饮食导致小鼠左心室心肌脂滴沉积明显增多[（13.8±0.86）/100 μm2比（4.7±0.51）/100 μm2，P<0.05]，而烟酰胺核糖可减轻高脂诱导的心肌纤维化[（6.2±0.77）/100 μm2比（13.8±0.86）/100 μm2，P=0.021]。免疫印迹显示脂毒性心肌组织中Sirt1及心肌肌浆网Ca2+-ATP酶（Serca2a）蛋白表达水平均较对照组降低（均P<0.05），Serca2a的乙酰化水平增高（均P<0.05），但烟酰胺核糖可增加Sirt1及Serca2a的表达并促进Serca2a的去乙酰化（均P<0.05）。 结论 烟酰胺核糖通过刺激Sirt1介导的Serca2a去乙酰化缓解高脂所致的脂毒性心肌病。 Objective To investigate the role and mechanism of nicotinamide riboside in obesity-related lipotoxic cardiomyopathy. Methods Male C57BL/6N mice at 8 weeks of age were randomly divided into the normal diet group (control group), high-fat diet group, and high-fat diet with nicotinamide ribose group (n=10 each). The mice on high-fat diet were modeled for obese mice lipotoxic cardiomyopathy over a24-week feeding. The mice in the high-fat diet with nicotinamide ribose group were administered with 40 mmol/L nicotinamide ribose in drinking water for 12 weeks on top of the high-fat diet. After modeling, all mice were measured for blood pressure of the caudal artery, collected of peripheral blood samples for testing of blood sugar, and lipids, and evaluated for heart function by echocardiography. Then the mice hearts were harvested and subjected to pathological and molecular biological studies. The heart weight and tibia length of the mice were measured by gross photography. The area of mice cardiomyocytes was quantified by staining with wheat germ agglutinin (WGA). Picrosirius red (PSR) staining was used to show the myocardial fibrosis in mice such that the effect of nicotinamide riboside on mice myocardial remodeling was evaluated. The lipid droplets infiltration in mice cardiomyocytes was examined by transmission electron microscopy. The protein expression levels of Sirt1 and Serca2a in mice cardiomyocytes were detected by Western blotting. Results Compared with the control group, the mice on high-fat diet presented significant increases in body weight, systolic blood pressure, diastolic blood pressure, total cholesterol, triglyceride and LDL-c levels (all P<0.05), and the elevation in these indicators indiced by high fat diet was alleviated by the addition of nicotinamide riboside (allP<0.05). The heart rate and blood sugar level in mice were not affected by high-fat diet with or without nicotinamide riboside. Compared with the control group, the mice on high-fat diet had higher or greater left ventricular ejection fraction and lower ratio of E to A peaks (E/A) on echocardiograpy (bothP<0.05), and addition of nicotinamide riboside increased the left ventricular ejection fraction and E/A ratio (bothP<0.05). Compared with the control group, the mice on high-fat diet had higher or greater ratio of heart weight to tibia length, cross-sectional area and collagen contents in cardiomyocytes of the left ventricle (allP<0.05), whereas addition of nicotinamide riboside was found to inhibit the myocardial hypertrophy and fibrosis as described above (allP<0.05). Compared with the control group, high-fat diet led to a significant increase in deposition of lipid droplets in the left ventricular myocardium[ (13.8±0.86) /100 μm2 vs (4.7±0.51) /100 μm 2, P<0.05], while nicotinamide ribose could alleviate such lipid-induced myocardial fibrosis[ (6.2±0.77) /100 μm2 vs (13.8±0.86) /100 μm 2, P=0.021]. Western blotting showed lower protein expression levels of Sirt1 in the lipotoxic myocardial tissues and sarco (endo) plasmic reticulum Ca2+-ATPase (Serca2a), along with increased acetylation of Serca2a in mice on high-fat diet compared with the control group (all P<0.05), while nicotinamide riboside was shown to increase Sirt1 and Serca2a expression and promote Serca2a deacetylation (allP<0.05) . Conclusion Nicotinamide riboside may alleviate high fat-induced lipotoxic cardiomyopathy by promoting Sirt1-mediated deacetylation of Serca2a.

    脂毒性心肌病烟酰胺核糖膳食,高脂心肌疾病,扩张型心内膜心肌纤维化症抗衰老酶1肌浆网钙转运ATP酶类去乙酰化

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    非诺贝特通过下调细胞周期蛋白D1表达诱导肺腺癌细胞周期阻滞

    王桂平李智斌周伟平张彦焘...
    34-40页
    查看更多>>摘要：目的 探讨非诺贝特（FEN）对肺腺癌A549细胞的增殖、细胞周期影响及其相关分子机制，为FEN治疗肺腺癌提供理论依据。 方法 实验设对照组[二甲基亚砜（DMSO）]和不同浓度FEN组（100、200、400 μmol/L），采用CCK-8法检测FEN对A549细胞增殖抑制率。将A549细胞分为对照组（DMSO）和100、200 μmol/L FEN组，通过流式细胞仪检测细胞周期；应用生物信息学技术揭示FEN抗肺腺癌分子机制，并筛选出治疗肺腺癌的核心靶点分子；应用实时荧光定量（qRT）-PCR及免疫印迹法验证FEN对核心靶点细胞周期蛋白D1（CCND1）基因及蛋白表达的影响。 结果 与对照组比较，随着FEN浓度增加，A549细胞增殖抑制率也逐渐增高（均P<0.05），且各FEN组增殖抑制率也随作用时间延长逐渐增高（均P<0.05）。FEN可诱导A549细胞产生G1期阻滞，100、200 μmol/L FEN组G1期细胞比率较对照组明显升高（均P<0.05）。生物信息学预测显示，FEN可能主要通过激动过氧化物酶增殖激活受体γ（PPARγ）和PPARα，调控CCND1等31个核心靶分子表达而发挥抗肺腺癌作用，而FEN对A549细胞G1期阻滞可能主要与CCND1的表达下调相关。qRT-PCR及免疫印迹验证实验也表明，与对照组比较，FEN可明显抑制CCND1基因及蛋白表达（均P<0.05）。 结论 FEN通过抑制CCND1分子表达而抑制A549细胞增殖，并将细胞周期阻滞在G1期。 Objective To investigate the effects of Fenofibrate (FEN) on proliferation and cell cycle of lung adenocarcinoma cell line A549 and its related molecular mechanism, so as to provide the theoretical rationale for FEN as a treatment of lung adenocarcinoma. Methods A control group (dimethyl sulfoxide, DMSO) and FEN groups at different doses (100, 200 and 400 μmol/L) were included in the study. The inhibitory rate of FEN on A549 cell proliferation was examined by CCK-8 assay. A549 cells were divided into the control group (DMSO) and 100, 200 μmol/L FEN groups, and the cell cycle was measured by flow cytometry. Bioinformatic technique was used to determine the molecular mechanism of FEN against lung adenocarcinoma, and the core target molecules for the treatment of lung adenocarcinoma were screened out. Real-time fluorescence quantitative (qRT) -PCR and Western blotting were used to verify the effect of FEN on the gene and protein expression of the core target cyclin D1 (CCND1) . Results Compared with the control group, the inhibition rate of A549 cell proliferation gradually increased along with higher concentrations of FEN (all P<0.05), and the inhibition rate of A549 cell proliferation in the FEN groups gradually increased with the time (allP<0.05). FEN could induce cell cycle arrest at G1 phase in A549 cells, and the proportions of G1 phase cells in 100 and 200μmol/L FEN groups were significantly elevated compared with that in the control group (allP<0.05). Bioinformatics prediction showed that FEN may play the role against lung adenocarcinoma mainly by activating PPARγ and PPARα receptors, and regulating the expressions of 31 core target molecules such as CCND1 the cell cycle arrest of A549 cells at G1 phase by FEN may be mainly related to down-regulation of CCND1 expression. qRT-PCR and Western blotting verified that FEN could significantly inhibit the gene and protein expression of CCND1 compared with that in the control group (all P<0.05) . Conclusion Fenofibrate may inhibit the proliferation of A549 cells by down-regulating CCND1 expression, and thereby arrest the cell cycle in G1 phase.

    肺腺癌非诺贝特细胞周期蛋白D1细胞周期细胞增殖计算生物学

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    CT引导下经皮肺穿刺活检术非诊断性结果的影响因素

    朱建彬屈耀铭王显龙汪洋...
    41-50页
    查看更多>>摘要：目的 评估CT引导下经皮肺穿刺活检术（PCTLB）非诊断性结果的相关影响因素，并探究每个非诊断类别最终恶性病变的发生率及风险。 方法 收集2014年1月至2017年9月于本院影像诊断科接受PCTLB的1028例的临床、影像学特征及病理诊断（包括穿刺诊断及最终诊断）资料。非诊断性结果分为非特异性良性病变、非典型细胞和正常肺组织或组织标本不足。通过单、多因素分析寻找与非诊断性结果及非诊断结果中恶性病变相关的影响因素。 结果 穿刺病理结果为非诊断性结果的比例为22.9%（235/1028），最终有28.1%（66/235）病例被诊断为恶性病变，在3类非诊断性病理类别中，来源于非特异性良性病变组为37.9%（25/66）、非典型细胞组34.8%（23/66）及正常肺组织或组织标本不足组27.3%（18/66）。最终恶性病变的风险比例在非典型细胞（88.5%）组最高。很短的穿刺标本长度（0~0.1 cm）（OR=4.606，P<0.05）及怀疑为良性病变（OR=14.597，P<0.05）是非诊断性结果的独立影响因素。在非诊断性结果中，实性病灶（OR=7.430，P<0.05）、术后发生高级别肺出血并发症（OR=6.411，P=0.003）、较长的穿刺标本长度（OR=0.141，P=0.003）、病理显示存在非典型细胞（OR=61.604，P<0.05）及正常肺组织或组织标本不足（OR=4.486，P=0.002）与最终诊断为恶性病变相关。 结论 在PCTLB的非诊断结果中，病理显示为非典型细胞、正常肺组织或组织标本不足与最终恶性诊断相关，并且强调被归类为非典型细胞组为恶性病变的可能性最高，对怀疑为恶性病变可能时，应进行重复的PCTLB或临床影像随访。 Objective To evaluate the influencing factors of non-diagnostic findings in percutaneous computed tomography-guided transthoracic lung biopsy (PCTLB), and the incidence and risk of eventual malignancy in each non-diagnostic category. Methods The clinical, imaging features and pathological diagnoses (including puncture biopsy and final diagnoses) of 1028 patients who underwent PCTLB in our Department of Imaging Diagnostics between January 2014 and September 2017 were retrieved. Non-diagnostic findings were classified as non-specific benign lesions, atypical cells, and normal lung tissue or inadequate tissue specimens. The influencing factors related to non-diagnostic findings and eventual malignancy with these non-diagnostic findings were determined via univariate and multivariate analyses. Results The proportion of non-diagnostic findings of puncture biopsy was 22.9% (235/1028). Of these, 28.1% (66/235) were eventually identified as malignancy. Among the three categories of non-diagnostic findings, 37.9% (25/66) were of benign lesions, 34.8% (23/66) were of atypical cells, and 27.3% (18/66) were of normal lung tissue or inadequate tissue specimens. The risk ratio for eventual malignancy was highest with the findings of atypical cells (88.5%). Very short length of puncture specimen (0-0.1 cm) (OR=4.606, P<0.05) and suspected benign lesions (OR=14.597, P<0.05) were independent influencing factors of non-diagnostic findings. Among the non-diagnostic findings, solid lesions (OR=7.430, P<0.05), post-procedural higher-grade hemorrhagic complications of the lung (OR=6.411, P=0.003), longer length of biopsy specimen (OR=0.141, P=0.003), presence of atypical cells identified by pathology (OR=61.604, P<0.05), and normal lung tissue or inadequate tissue specimens (OR=4.486, P=0.002) were associated with eventual diagnosis of malignancy. Conclusion Among the non-diagnostic findings of PCTLB, pathological identification of atypical cells, normal lung tissue or inadequate tissue specimens were associated with eventual diagnosis of malignancy. It should be emphasised that atypical cells as a category of non-diagnostic findings are linked to the highest likelihood of malignancy. Repeat PCTLB or follow-up on clinical imaging should be performed when malignant lesions are suspected.

    体层摄影术,螺旋计算机经皮肺穿刺活检术非诊断结果非典型细胞影响因素

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    黄连素降低耐环丙沙星的铜绿假单胞菌耐药性及对耐药基因GyrA的影响

    韩菊梅肖英伦刘新艳
    51-56页
    查看更多>>摘要：目的 观察黄连素应用后铜绿假单胞菌（PA）对环丙沙星的最低抑菌浓度（MIC）的变化和PA菌耐药基因GyrA突变情况。 方法 收集2021年1月至10月临床分离耐环丙沙星铜绿假单胞菌25株，测定单用或联用环丙沙星和黄连素的最低抑菌浓度（MIC），计算抑菌浓度指数（FIC），筛选出联用呈协同作用菌株，并和原临床分离耐药菌配对比较。PCR扩增PA主要耐喹诺酮的GyrA基因，并对PCR扩增产物进行测序分析。 结果 单用环丙沙星时，25株PA对环丙沙星的MIC均≥8 mg/L，呈耐药。单用黄连素时，MIC值为0.25~4 g/L。环丙沙星联合黄连素浓度0.125~1 g/L后，PA对环丙沙星的MIC值均有不同程度下降，下降率为100%，联用呈协同作用菌株为8株，占比32%,FIC≤0.5。联合黄连素后，黄连素应用后呈协同作用的PA菌与黄连素应用前PA菌均获得367 bp的目的片段，均有GyrA基因的表达，未发现GyrA基因突变。 结论 黄连素能增加环丙沙星对PA的抗菌作用，降低临床耐药PA菌的耐药性。其耐药性降低，并未引起耐药基因GyrA基因的突变逆转。 Objective To determine the changes in minimum inhibitory concentration (MIC) of Ciprofloxacin against Pseudomonas aeruginosa (PA) and the mutation in drug-resistant gene GyrA after use of Berberine. Methods Twenty-five strains of Ciprofloxacin-resistant PA clinically isolated between January 2021 and October 2021 were included in the study. The MIC against PA with Ciprofloxacin or Berberine alone or in combination was determined, respectively, and thereby the fractional inhibitory concentration (FIC) index was calculated. We sorted out the strains with synergistic response to Ciprofloxacin and Berberine combination and compared them with the original stains of drug-resistant PA. The major gene of Quinolone-resistance in PA, GyrA, was amplified by PCR, and subjected to sequencing analysis of the PCR amplification products. Results When Ciprofloxacin was used alone, its MIC against these 25 PA strains was ≥8 mg/L, indicating drug resistance. When Berberine was used alone, its MIC was 0.25-4 g/L. When used in combination with Berberine at doses of 0.125-1 g/L, the MIC of Ciprofloxacin against PA showed varying decrease in 100% of the strains. There were 8 strains (32%) showing synergistic response to the combination treatment, with a Ciprofloxacin FIC ≤ 0.5. A 367 bp target fragment was obtained from PA strains that showed synergistic response to Ciprofloxacin-Berberine combination and those without Berberine use. Either strain was expressing the GyrA gene with no mutations noted. Conclusion Berberine may increase the antibacterial effect of Ciprofloxacin on PA and reduce the drug resistance of clinical drug-resistant PA. Such attenuation in the drug resistance does not lead to mutational reversion of the drug-resistant gene GyrA.

    铜绿假单胞菌小檗碱环丙沙星GyrA基因微生物敏感性试验

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    运用CRISPR-dCas9-VP64基因编辑技术验证microRNA-140的独立转录启动位点

    郝耀陈丽韩永斌原野...
    57-64页
    查看更多>>摘要：目的 探讨microRNA-140（miR-140）是否有独立于其宿主基因WWP2的转录启动子，从而为骨性关节炎的治疗提供新的思路。 方法 通过共转染CRISPR-dCas9-VP64和ACAN gRNA慢病毒的SW1353细胞验证软骨细胞特征性基因ACAN的转录水平。通过构建能稳定表达CRISPR-dCas9-VP64的SW1353细胞并结合多条sgRNA（single guide RNA, sgRNA/gRNA）、人类髋关节软骨RNA测序结果验证miR-140是否与WWP2基因共表达，是否有独立的转录启动位点。 结果 共转染CRISPR-dCas9-VP64和ACAN gRNA慢病毒的SW1353细胞能显著提高软骨细胞特征性基因ACAN的转录水平。从基因水平、蛋白水平证实成功构建能稳定表达CRISPR-dCas9-VP64的SW1353细胞。通过人类髋关节软骨RNA测序结果设计多条WWP2 gRNA以及miR-140启动子gRNA。实时定量PCR结果显示miR-140的表达既受其宿主基因WWP2调控，同时也有其独立的转录位点。 结论 运用CRISPR-Cas9基因编辑技术成功构建稳定表达CRISPR-dCas9-VP64蛋白的SW1353细胞能极大减少实验中样本间的差异化，保证实验结果的一致性。结合多条gRNA证明了miR-140既与WWP2共表达，同时也受其独立的转录启动子调控转录。运用CRISPR-dCas9-VP64d-gRNA能单独调控miR-140的表达，同时其宿主基因的表达不受影响，有潜力作为新兴的基因治疗方法治疗骨性关节炎。 Objective To explore whether microRNA-140 (miR-140) has a transcriptional promoter independent of its host gene WWP2, so as to provide new ideas for the treatment of osteoarthritis. Methods The transcript level of chondrocyte characteristic gene ACAN was verified by co-transfecting SW1353 cells with CRISPR-dCas9-VP64 and ACAN gRNA lentivirus. By constructing SW1353 cells that can stably express CRISPR-dCas9-VP64 and combining multiple sgRNA (single guide RNA, sgRNA/gRNA) and human hip cartilage RNA sequencing results to verify whether miR-140 is co-expressed with WWP2 gene, whether there is an independent Transcription initiation site. Results SW1353 cells co-transfected with CRISPR-dCas9-VP64 and ACAN gRNA lentivirus could significantly increase the transcription level of chondrocyte characteristic gene ACAN. The successful construction of SW1353 cells that can stably express CRISPR-dCas9-VP64 was confirmed from the gene level and protein level. Multiple WWP2 gRNAs and miR-140 promoter gRNAs were designed based on the RNA sequencing results of human hip cartilage. The results of real-time quantitative PCR showed that the expression of miR-140 was not only regulated by its host gene WWP2, but also had its independent transcription site. Conclusions Using CRISPR-Cas9 gene editing technology to successfully construct SW1353 cells stably expressing CRISPR-dCas9-VP64 protein can greatly reduce the difference between samples in the experiment and ensure the consistency of the experimental results. Combining multiple gRNAs proved that miR-140 was not only co-expressed with WWP2, but also regulated by its independent transcriptional promoter. The use of CRISPR-dCas9-VP64d-gRNA can regulate the expression of miR-140 alone, while the expression of its host gene is not affected, which has the potential to be used as an emerging gene therapy method for the treatment of osteoarthritis.

    CRISPR-dCas9-VP64骨性关节炎非编码小RNA-140含WW域E3泛素蛋白连接酶2

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    外泌体介导circ_0000218靶向微RNA-144调控胆管癌细胞的增殖、迁移和侵袭

    渠海孙衍赵立军张宁...
    65-71页
    查看更多>>摘要：目的 探讨外泌体介导的circ_0000218对胆管癌细胞增殖、迁移和侵袭的影响及其分子机制。 方法 分离提取胆管癌细胞外泌体，qRT-PCR检测外泌体中circ_0000218表达水平，将si-circ_0000218转染至胆管癌细胞，提取外泌体分别与RBE和HuH-28细胞共培养；CCK8检测细胞活力；Transwell检测细胞迁移和侵袭；免疫印迹检测相关蛋白的表达；双荧光素酶报告基因实验检测circ_0000218和miR-144的靶向关系；功能回复实验观察circ_0000218/miR-144轴对CCA细胞增殖、迁移和侵袭的影响。 结果 circ_0000218在胆管癌组织和细胞中高表达（P<0.05）；外泌体介导的敲低circ_0000218可降低RBE和HuH-28细胞活力、迁移和侵袭数目以及Ki67、MMP-2、MMP-9表达水平（P<0.05）；circ_0000218靶向调控miR-144表达（P<0.05），且下调miR-144逆转了敲低circ_0000218表达对RBE、HuH-28细胞的增殖、迁移和侵袭的影响（P<0.05）。 结论 外泌体介导的circ_0000218可通过靶向调控miR-144表达影响胆管癌细胞的增殖、迁移和侵袭。 Objective To investigate the effect of exosome-mediated circ_0000218 on the proliferation, migration and invasion of cholangiocarcinoma cells and its molecular mechanism. Methods The exosomes were isolated and extracted from cholangiocarcinoma cells, and the expression level of circ_0000218 in the exosomes was determined by qRT-PCR. si-circ_0000218 was transfected into cholangiocarcinoma cells, and extracted exosomes were co-cultured with RBE and Huh-28 cells, respectively. Cell counting kit-8 (CCK8) assay was used to examine cell viability. Transwell assay was used to detect cell migration and invasion. Western blotting was used to measure the expression levels of related proteins. Dual luciferase reporter assay was used to identify the targeting relationship between circ_0000218 and miR-144. Functional recovery assay was used to detect the effects of circ_0000218/miR-144 axis on the proliferation, migration and invasion of CCA cells. Results circ_0000218 was highly expressed in cholangiocarcinoma tissues and cells (P<0.05). Exosome-mediated knockdown of circ_0000218 reduced the RBE and Huh-28 cell viability, migration and invasion numbers, and decreased the expression levels of Ki67, MMP-2 and MMP-9 (allP<0.05). circ_0000218 targeted and regulated the expression of miR-144 (P<0.05), and the down-regulation of miR-144 reversed the effects of knockdown of circ_0000218 expression on the proliferation, migration and invasion of RBE and Huh-28 cells (allP<0.05) . Conclusion Exosome-mediated circ_0000218 may affect proliferation, migration and invasion of cholangiocarcinoma cells by targeting and regulating the expression of miR-144.

    胆管癌外泌体环状RNA增殖迁移侵袭

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    神经根阻滞联合椎间孔镜治疗腰椎间盘突出症的疗效观察

    徐云杉张锦洪马双双夏绍祥...
    72-76页
    查看更多>>摘要：探讨选择性神经根阻滞术（SNRB）联合侧路经皮椎间孔镜下髓核摘除术（PTED）在治疗多节段腰椎间盘突出中的意义。研究发现选择性神经根阻滞可为多节段腰椎间盘突出症患者提供定位诊断，联合PTED术可实现精准减压，可取得良好的治疗效果。 To explore the clinical efficacy of selective nerve root block (SNRB) in percutaneous transforaminal endoscopy discectomy (PTED) for multi-segmental lumbar disc herniation (MLDH). SNRB was effective for diagnosing MLDH, which could provide reliable evidence for PTED to accomplish precise decompression and minimally invasive surgery.

    腰椎间盘突出症多节段选择性神经根阻滞经皮椎间孔镜髓核摘除术

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