长链非编码RNA反义RNA 1靶向微小RNA-501-3p对食管癌细胞恶性生物学行为影响的实验研究

目的探讨长链非编码RNA(lncRNA)胸腺生成素反义RNA 1(TMPO-AS1)对食管癌细胞恶性生物学行为的影响及其机制。方法2016年6月至2019年12月,实时荧光定量聚合酶链反应(RT-qPCR)检测20例食管癌组织和癌旁组织中lncRNA TMPO-AS1和微小RNA(miR)-501-3p的水平,食管癌Eca109细胞分为lncRNA TMPO-AS1干扰组(转染si-NC和si-TMPO-AS1);miR-501-3p过表达组(转染miR-NC和miR-501-3p);lncRNA TMPO-AS1过表达组(转染pcDNA和pcDNA-TMPO-AS1);lncRNA TMPO-AS1和miR-501-3p共抑制组(转染si-TMPO-AS1+anti-miR-NC和si-TMPO-AS1+anti-miR-501-3p)。噻唑蓝(MTT)法、流式细胞术和Transwell实验分别检测食管癌Eca109细胞增殖、凋亡率、细胞迁移和侵袭能力,蛋白质印迹法(Western blot)检测相关蛋白水平,双荧光素酶报告系统验证lncRNA TMPO-AS1与miR-501-3p的调控关系。两组间比较采用独立样本t检验,多组间比较采用单因素方差进行分析。结果食管癌组织组中的lncRNA TMPO-AS1水平(2.64±0.26)高于癌旁组织(1.00±0.09,t=26.657,P<0.05),miR-501-3p水平(0.44±0.04)低于癌旁组织(1.00±0.07,t=31.063,P<0.05);干扰lncRNA TMPO-AS1的Eca109细胞24 h增殖(0.36±0.03)低于si-NC组(0.39±0.03,t=2.121,P<0.05),48 h增殖(0.45±0.04)低于si-NC组(0.75±0.07,t=11.163,P<0.05),72 h增殖(0.57±0.05)低于si-NC组(1.16±0.09,t=17.191,P<0.05),迁移细胞数[(54.14±5.65)个]低于si-NC组[(113.02±9.87)个,t=15.531,P<0.05],侵袭细胞数[(47.69±5.01)个]低于si-NC组[(96.32±9.88)个,t=13.169,P<0.05],细胞凋亡率[(22.15±2.22)%]高于si-NC组[(6.98±0.69)%,t=19.576,P<0.05];过表达miR-501-3p的Eca109细胞48 h增殖(0.51±0.05)低于miR-NC组(0.77±0.07,t=9.067,P<0.05),72 h增殖(0.65±0.06)低于miR-NC组(1.15±0.09,t=13.867,P<0.05),迁移细胞数[(61.23±6.33)个]低于miR-NC组[(115.25±9.25)个,t=14.458,P<0.05],侵袭细胞数[(53.14±5.33)]低于miR-NC组[(98.32±8.47)个,t=13.543,P<0.05],细胞凋亡[(19.58±1.74)%]高于miR-NC组[(7.23±0.77)%,t=19.471,P<0.05],差异均有统计学意义。结论lncRNA TMPO-AS1通过靶向miR-501-3p促进食管癌细胞的恶性生物学行为,可被视为食管癌患者的潜在治疗靶标。     

腹腔注射舒芬太尼对SD大鼠气道反应性的影响

目的观察腹腔注射舒芬太尼对SD大鼠气道反应性的影响。方法将32只雄性SD大鼠按随机数字表法分为4组,正常对照为A组(n=8,腹腔注射生理盐水,3 ml);实验组,B1组(n=8,腹腔注射舒芬太尼60μg/kg,3 ml)、B2组(n=8,腹腔注射舒芬太尼90μg/kg,3 ml)和B3组(n=8,腹腔注射舒芬太尼120μg/kg,3 ml)。第1天对4组大鼠进行气道阻力和气道反应性应用非侵入性气道阻力仪测定并作为参考基线。第15天按照分组要求对4组大鼠进行腹腔注射干预措施,测定腹腔注射后1、2、3、5、10 min气道阻力(specific airway resistances,Raw)(cmH2O·s),第30天按照分组要求对4组大鼠进行腹腔注射干预措施,3 min后进行气道反应性测定。采用t检验和Fisher确切概率法对实验数据进行分析。结果实验第1天,与对照组A组比较,B1组、B2组、B3组大鼠腹腔注射前基线气道阻力(3.27±0.74比3.35±0.65、3.30±0.81比3.35±0.65、3.41±0.68比3.35±0.65)差异无统计学意义(t=0.230,P>0.05、t=0.136,P>0.05、t=0.180,P>0.05);实验第15天,与对照组A组干预后1、2、3、5、10 min的气道阻力比较,B1组在1、2 min气道阻力(4.15±1.27比3.95±0.89、4.74±1.05比3.78±0.85)差异无统计学意义(t=0.365,P>0.05、t=2.010,P>0.05),在3、5 min气道阻力(4.92±1.33比3.55±0.69、4.89±1.43比3.47±0.71)差异有统计学意义(t=2.586,P<0.05、t=2.516,P<0.05),在10 min气道阻力(3.86±1.28比3.41±0.69)差异无统计学意义(t=0.875,P>0.05);B2组在1 min气道阻力(4.77±1.15比3.95±0.89)差异无统计学意义(t=1.595,P>0.05),在2、3、5 min气道阻力(6.01±1.47比3.78±0.85、6.95±1.68比3.55±0.69、5.75±1.52比3.47±0.71)差异有统计学意义(t=3.715,P<0.05、t=5.295,P<0.05、t=3.844,P<0.05),在10 min气道阻力(3.95±1.18比3.41±0.69)差异无统计学意义(t=1.117,P>0.05);B3组在1、2、3、5 min气道阻力(5.14±1.25比3.95±0.89、7.91±1.35比3.78±0.85、8.15±1.48比3.55±0.69、7.69±1.23比3.47±0.71)差异有统计学意义(t=2.194,P<0.05、t=7.322,P<0.05、t=7.968,P<0.05、t=8.404,P<0.05),在10 min气道阻力(4.16±1.08比3.41±0.69)差异无统计学意义(t=1.655,P>0.05);实验第30天,4组大鼠腹腔注射后气道反应性阳性率为,与对照组A组(0%)比较,B1组37.5%差异无统计学意义(P>0.05)、B2组75%差异有统计学意义(P<0.05)和B3组87.5%差异有统计学意义(P<0.05)。结论腹腔注射舒芬太尼对大鼠呼吸系统的影响表现在气道阻力增加和气道反应性增高,并呈剂量依赖性。但这种变化在短时间内逐渐增加并达到高峰后,进而逐渐减弱。     

长链非编码RNA Linc00472靶向微小RNA-381对骨肉瘤细胞增殖、凋亡、迁移、侵袭的影响

目的观察长链非编码RNA Linc00472(LncRNA Linc00472)靶向调控微小RNA-381(miR-381)对骨肉瘤细胞增殖、凋亡、迁移、侵袭的影响。方法收集2017年3月至2018年5月郑州大学第一附属医院收治的骨肉瘤患者29例为研究对象,实时荧光定量聚合酶链反应(RT-qPCR)检测骨肉瘤组织和瘤旁组织中Linc00472的表达。将骨肉瘤U2OS细胞分为pcDNA3.1组、pcDNA3.1-Linc00472组、pcDNA3.1-Linc00472+miR-NC组、pcDNA3.1-Linc00472+miR-381组。甲基噻唑基四唑(MTT)检测细胞增殖;流式细胞术检测细胞凋亡;Transwell实验检测细胞迁移及侵袭。双荧光素酶报告实验鉴定Linc00472与miR-381的靶向关系。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析。结果Linc00472在骨肉瘤组织中的表达水平低于瘤旁组织(0.31±0.03比1.03±0.10,t=37.138,P<0.05),差异有统计学意义;与pcDNA3.1组比较,pcDNA3.1-Linc00472组细胞存活率[(100.29±10.12)%比(53.29±5.44)%]低于pcDNA3.1组(t=12.272,P<0.05),差异有统计学意义,迁移细胞数[(266.00±23.61)个比(124.00±12.01)个]与侵袭细胞数[(131.00±13.06)个比(62.00±6.24)个]少于pcDNA3.1组(t=16.082,P<0.05;t=14.301,P<0.05),差异有统计学意义,而细胞凋亡率[(8.58±0.91)%比(26.61±2.64)%]高于pcDNA3.1组(t=19.370,P<0.05),差异有统计学意义;miR-381过表达可抑制野生型载体WT-Linc00472细胞的荧光素酶活性(t=19.827,P<0.05),差异有统计学意义;与pcDNA3.1-Linc00472+miR-NC组比较,pcDNA3.1-Linc00472+miR-381组可增强细胞增殖[(53.17±5.33)%比(85.26±8.62)%]、迁移[(122.00±12.31)个比(223.00±22.18)个]及侵袭[(64.00±6.36)个比(104.00±10.20)个]能力高于pcDNA3.1-Linc00472+miR-NC组(t=9.499,P<0.05;t=11.945,P<0.05;t=9.983,P<0.05),细胞凋亡率[(26.11±2.62)%比(13.11±1.34)%]低于pcDNA3.1-Linc00472+miR-NC组(t=13.253,P<0.05),差异有统计学意义。结论LncRNA Linc00472通过靶向调控miR-381可抑制骨肉瘤细胞增殖、迁移及侵袭并诱导细胞凋亡。     

腹腔镜结直肠癌手术中转开腹列线图预测模型的构建

目的分析腹腔镜结直肠癌手术发生中转开腹的危险因素,建立列线图预测模型指导临床决策,并分析其短期预后影响。方法收集2017年5月至2020年5月西安交通大学第一附属医院行腹腔镜手术的764例结直肠癌患者的病例资料,分为中转开腹组(60例)及腹腔镜组(704例)。采用Logistic回归分析中转开腹的独立危险因素,并用R语言构建中转开腹列线图预测模型。通过受试者工作特征曲线(ROC)下面积评估列线图预测模型效能。通过绘制校准图进行一致性检验。并比较两组患者术中及术后恢复情况。结果本研究共纳入患者764例,其中60例发生中转开腹,中转开腹率为7.9%(60/764)。肿瘤位置位于直肠[比值比(OR)=1.846,P<0.05)]、美国麻醉医生协会(ASA)评分Ⅲ、Ⅳ级(OR=2.381,P<0.05)、腹部手术史(OR=3.652,P<0.01)、肿瘤长径≥5 cm(OR=2.704,P<0.05)和体重指数(OR=1.109,P<0.05)是发生中转开腹的独立危险因素,上述数据差异均有统计学意义。成功构建列线图预测模型,模型ROC曲线下面积为0.794。校正曲线显示预测结果与实际结果有较好的重合度。中转开腹组手术时间高于腹腔镜组(210 min比190 min,Z=-2.670,P<0.05);中转开腹组出血量高于腹腔镜组(170 ml比120 ml,Z=-6.018,P<0.01);中转开腹组术后住院天数高于腹腔镜组(10 d比9 d,Z=-2.134,P<0.05)。中转开腹组肠梗阻发生率高于腹腔镜组[10.0%(6/60)比4.3%(15/704),χ²=4.055,P<0.05];伤口感染率高于腹腔镜组[15.0%(9/60)比4.7%(33/704),χ²=11.318,P<0.05],上述数据差异均有统计学意义。结论肿瘤位置位于直肠、ASA评分Ⅲ、Ⅳ级、腹部手术史、肿瘤长径≥5 cm和体重指数是腹腔镜结直肠癌手术发生中转开腹的独立危险因素,据此建立的预测模型具有可靠的预测能力。     

肩关节上关节囊重建治疗肩袖损伤适应证和移植物处理方法研究进展

目的对肩关节上关节囊重建治疗肩袖损伤适应证和移植物处理方法的研究进展进行综述。方法广泛查阅近年来国内外有关肩关节上关节囊重建治疗肩袖损伤的文献，并进行总结分析。结果肩关节上关节囊重建可以有效恢复肩关节上方稳定性，其手术适应证包括不可修复巨大肩袖撕裂、巨大肩袖撕裂合并肩关节假性麻痹、肩袖分层撕裂以及中或大的肩袖撕裂合并肩袖严重退变。为了获得较好的腱骨愈合以及降低移植物远期再撕裂率，必须选择合适厚度的移植物，并以适当的强度固定移植物并恢复其连续性。结论肩关节上关节囊重建的手术应用已发展至上关节囊加强重建，手术适应证从肩袖实质性巨大缺损扩展至肩袖严重退变。移植物处理是上关节囊重建手术成功的关键。     

锌指蛋白 A20 对兔腰椎间盘退变影响的实验研究

目的探讨锌指蛋白 A20 对兔腰椎间盘退变的影响。方法取 3 月龄新西兰大白兔 26 只，体质量 2.0～2.5 kg，经腹细针穿刺法制备 L_3、4、L_4、5、L_5、6 椎间盘退变模型，其中 24 只术后 4 周 MRI 检查明确造模成功，随机分为 4 组（n=6），于目标椎间盘中分别注射锌指蛋白 A20 过表达腺病毒（过表达 A20 组）、空载体腺病毒（空载体组）、PBS 液（对照组）、锌指蛋白 A20干扰腺病毒（干扰 A20 组）。于注射前 1 d 及注射后 1、2、3、6 d 行生物反应综合评分；注射后 2、4、8 周，各组行 MRI 检查并测量 T2 弛豫时间（T2 信号值）后，取材行阿利辛蓝染色观察椎间盘髓核细胞退变情况，免疫组织化学染色检测锌指蛋白 A20 以及椎间盘退变相关指标（Ⅱ型胶原、蛋白聚糖）的表达，Western blot 检测锌指蛋白 A20、NF-κB 结合蛋白［P65、磷酸化 P65（phosphate P65，P-P65）、Ⅱ型胶原、蛋白聚糖］、自噬相关蛋白［LC3 （LC3Ⅱ/LC3Ⅰ）、P62］以及炎症因子（TNF-α、IL-1β）的表达。 结果各组注射后各时间点生物反应综合评分均明显低于注射前 1 d（P<0.05）；注射后 6 d 干扰 A20 组评分明显低于其他组（P<0.05），其他组间比较差异均无统计学意义（P>0.05）。MRI 检测提示，注射后 2、4、8 周过表达 A20 组 T2 信号值均最高（P<0.05），2、4 周时干扰 A20 组最低（P<0.05），其余组间差异均无统计学意义（P>0.05）。阿利辛蓝染色显示，注射后 4 周过表达 A20 组蛋白聚糖含量最高（P<0.05）、干扰 A20 组最低（P<0.05）；8 周时过表达 A20 组蛋白聚糖含量显著高于其他组（P<0.05），其他组间比较差异无统计学意义（P>0.05）。免疫组织化学染色示，锌指蛋白 A20、Ⅱ型胶原、蛋白聚糖表达过表达 A20 组最高（P<0.05），干扰 A20 组上述蛋白表达最低（P<0.05）。Western blot 检测示锌指蛋白 A20、蛋白聚糖、Ⅱ型胶原、LC3 （LC3Ⅱ/LC3Ⅰ）蛋白相对表达量过表达 A20 组最高、干扰 A20 组最低，而 P-P65、TNF-α、IL-1β、P62 蛋白相对表达量过表达 A20 组最低、干扰 A20 组最高，与其他组比较差异均有统计学意义（P<0.05）；各组 P65 蛋白相对表达量差异均无统计学意义（P>0.05）。 结论锌指蛋白 A20 能通过抑制炎症反应，有效延缓兔腰椎间盘退变的进程。     

基于电子鼻技术的不同品质鹿茸饮片气味特征分析

目的:基于电子鼻技术对不同品质鹿茸饮片气味特征进行分析与表征。方法:采用PEN3型电子鼻系统,分析22批鹿茸样品的气味特征,以对传感器响应值为指标,进行传感器区别贡献率分析(Loadings)、主成分分析(PCA)、线性判别分析(LDA)。结果:Loadings分析表明,5个传感器对鹿茸饮片气味特征具有较好的响应,不同规格鹿茸饮片气味差异贡献率主要体现为氮氧化物类、甲烷等短链烷烃、有机硫化物、醇醚醛酮类、芳香成分、无机硫化物等成分;PCA表明不同品种与规格鹿茸饮片,其气味的差异性比较明显;LDA发现不同品种及规格鹿茸饮片(除梅花鹿白片与粉片外)样品的气味差异均较明显,表明构成气味的物质存在差异性。结论:电子鼻技术可阐明不同品质鹿茸饮片气味的物质基础;不同品质鹿茸饮片气味存在差异性可揭示鹿茸饮片气味的科学内涵并为其质量控制提供参考。     

Closed reduction of glenohumeral dislocations with associated tuberosity fracture in the emergency department is safe

AimA fracture of the tuberosity is associated with 16% of glenohumeral dislocations. Extension of the fracture into the humeral neck can occur during closed manipulation, leading some to suggest that all such injures should be managed under general anaesthesia in the operating theatre. The purpose of this study was to establish the safety of reduction of glenohumeral dislocations with tuberosity fractures in the emergency department (ED).Patients and methodsWe reviewed 188 consecutive glenohumeral dislocations with associated tuberosity fractures identified from a prospective orthopaedic trauma database. Patient demographics, injury details, emergency department management and complications were recorded. The method of reduction, sedation, grade of clinician and outcome were documented.ResultsThe mean age was 61 years (range 18–96 years) with 79 males and 109 females. The majority of injuries (146, 78%) occurred after a fall from standing height. Closed reduction under sedation in the ED was successful in 162 (86%) cases. Of the remainder, 22 (11%) failed closed reduction under sedation and subsequently went to theatre and 6 (3%) were deemed not suitable for ED manipulation. At presentation 35 (19%) patients had a nerve injury, of which 29 (90%) resolved spontaneously. Two iatrogenic fractures occurred during close manipulation, one in the ED and the other in the operating theatre. Therefore, the risk of iatrogenic propagation of the fracture into the proximal humerus neck was 0.5% if the reduction was performed in the ED, and 1% over-all. More than two attempted reductions predicted a failed ED reduction (P = 0.001).ConclusionClosed reduction of glenohumeral dislocations with associated tuberosity fractures in the ED is safe, with a rate of iatrogenic fracture of 1%. These injuries should be managed by those with appropriate experience only after two adequate radiographic views. In cases where there is ambiguity over the integrity of the humeral neck, reduction should be delayed until multiplanar CT imaging has been obtained.     

Outcomes following quadriceps tendon repair using transosseous tunnels versus suture anchors: A systematic review

IntroductionTwo major techniques are used to repair complete quadriceps tendon ruptures, transosseous tunnel (TT) and the suture anchor (SA). There are multiple studies comparing the biomechanical outcomes of repairs performed with TT or SA. Our purpose was to compare the clinical outcomes following quadriceps tendon repair using SA and TT fixation techniques.MethodsThree major search engines were used with predetermined keyword searches to perform a systematic review of literature. These studies were independently scanned by two reviewers using PRISMA criterion. All included studies had to include at least one of the following outcome measures: range of motion (ROM), Lysholm score, complications, and/or re-ruptures.ResultsUsing three major search engines, 1039 articles were identified. After removing duplicates and screening for inclusion, 49 articles were reviewed. Two independent reviewers searched the studies to meet the inclusion criteria, and eight studies were selected. These eight studies included 156 knees in the TT group and 54 knees in the SA group. The TT group had a significantly better ROM after QT repair (132.5° versus 127.0°, p = 0.02). There was no significant difference in Lysholm scores between the TT group (92.6) and SA group (91.0, p = 0.11). There were significantly more complications in SA groups (9.3% versus 1.3%, p = 0.013), but not a significant difference in re-rupture rate between those undergoing SA vs. TT repair (3.7% versus 0%, p = 0.065). The SA group had a significantly higher age at time of surgery (63.62 vs. 54.32)ConclusionThe current study suggests that, following quadriceps tendon rupture, there are no significant differences in functional outcome between TT and SA techniques. Those undergoing TT repair attained a statistically significantly greater final ROM but this difference may not be clinically relevant. There was a statistically significantly higher rate of post-operative complications using SA technique.