目的 探讨心肌细胞RyR2和L型钙通道的基因变异与室性心律失常和心源性猝死的相关性。方法 回顾分析2010年1月—2012年12月在我院就诊的慢性心力衰竭患者622例的临床资料,并选取同一时期体检中心体检的健康人群516例作为对照组,门诊或者电话随访记录慢性心力衰竭患者的死亡为终点,通过候选基因分析可能具有相关功能的4个基因变异,rs41315858(G1885E)、rs3766871(G1886S)、rs790896(G>A)和rs723672(T>C),采用Logestic、Cox回归分析对4个候选基因变异进行相关性研究。结果 入选622例慢性心力衰竭患者和516例对照组,基因分析结果显示RyR2上的基因变异rs376687lA等位基因携带可以增加慢性心力衰竭患者发生室性心律失常的风险性;校正可能与该疾病相关的危险因素后,rs376687lA等位基因携带会增加心源性死亡和心源性猝死的风险,RyR2上的基因变异rs790896A等位基因携带可以降低心源性猝死风险。结论 RyR2上的基因变异rs376687lA是室性心律失常和心源性猝死的遗传学预测因子,而rs790896A等位基因是慢性心力衰竭患者的保护因子,可降低室性心律失常和心源性猝死的风险。

Objective To investigate the myocardial cells RyR2 and L-type calcium channel gene variants with ventricular arrhythmias and sudden cardiac death correlation. Methods Retrospective analysis of patients with chronic heart failure from January 2010 to December 2012 in our hospital including 622 cases of clinical data, and to select 516 cases of healthy people in medical examination center during the same period as a control group.Clinic or telephone follow-up recorded chronic patients with heart failure and sudden death acting as end. We analyzed possible candidate genes, according to four gene variants related functions, rs41315858 (G1885E), rs3766871 (G1886S), rs790896 (G> A) and rs723672 (T> C), by using Logestic, Cox regression analysis of four candidate gene variants for related research. Results 622 cases of chronic heart failure patients were enrolled and 516 patients in the control group. Genetic analysis showed that the gene variant alleles carried rs376687lA RyR2 may increase in patients with chronic heart failure ventricular arrhythmia risk; correction may be associated with the disease after risk factors, rs376687lA allele carries an increased risk of cardiogenic death and sudden cardiac death, and gene mutation alleles carried on rs790896A RyR2 can reduce the risk of sudden cardiac death. Conclusion Gene mutation rs376687lA RyR2 on genetics is predictor of ventricular arrhythmias and sudden cardiac death, and rs790896A allele is protective factor in patients with chronic heart failure which can be reduced ventricular arrhythmias and sudden cardiac death in risk.

神经元特异性烯醇化酶(neuron-specific enolase, NSE)是糖酵解途径中一种重要的同工酶;特异性位于神经元和神经内分泌细胞中。小细胞肺癌(small cell lung cancer,SCLC)细胞浆内含有神经内分泌颗粒,具有神经内分泌分化的特征,为恶性程度高的神经内分泌系统肿瘤。因此,NSE是SCLC诊断中最敏感的肿瘤标志物,在SCLC的临床诊断、治疗、预后均有重要应用价值,科学合理联合检测肿瘤标记物,将能为临床诊疗工作提供有力的帮助。

目的 回顾性分析支气管肺泡细胞癌(bronchioloalveolar carcinoma,BAC)临床特点,提高早期BAC的确诊率,减少误诊。方法 对2013年—2014年间在我院确诊的BAC病例5例的临床资料进行回顾性分析。结果 5例患者中男4例,女1例,年龄在50～73岁之间,在社区医院均曾误诊为肺炎,所有患者均在我院经支气管镜肺活检后确诊为BAC。结论 BAC是一种较为特殊的肺癌,临床上症状无特异性,极易误诊为普通肺炎。由于BAC预后差,误诊后果严重,对初诊为普通肺炎的患者经常规抗感染治疗后临床症状及影像学表现改善不明显时,应及时进行各项检查、明确诊断,以便有效改善患者预后。

Objective To elevate the clinical diagnosis of BAC (bronchioloalveolar carcinoma) so as to reduce misdiagnosis by using retrospective analyses. Methods Retrospective analyses were used to study the five BAC patients, who were diagnosed in our hospital from 2013 to 2014. Results 4 of 5 male, and 1 female, age between 50 and 73, were diagnosed as pneumonia. All of them were made a definite diagnosis as BAC after performing transbronchial lung biopsy. Conclusion BAC is an exceptional lung adenocarcinoma and there is no specific clinic symptom. BAC was easily misdiagnosed as common lung pneumonia. There will be serious consequences after misdiagnosis of BAC due to its poor prognosis. Those patients who were misdiagnosed as common lung disease, but there was no obvious improvement after accepting long anti-infective therapy and there was negatively detection of pathogenic bacteria in them, are needed to perform all other clinical examination to clarify a diagnosis, and to further improve the prognosis of the patients.

目的 探讨磷酸二酯酶4抑制剂对人肺泡巨噬细胞(AM)吞噬非生物性颗粒及革兰阳性菌、阴性菌能力的影响。方法 使用Ficolll-Hypaque密度梯度法将外周血单核细胞分离的静脉血,在含有2 ng/m GM-CSF的培养液中经12天诱导培养成AM替代细胞模型—单核细胞源性巨噬细胞(MDM)。用酶标仪检测MDM经磷酸二酯酶4抑制剂Rolipram预处理过夜(16~18 h)后吞噬荧光标记的非生物颗粒Beads和热灭活的流感嗜血杆菌(H.influenzae)、金黄色葡萄球菌(S.aureus)量的改变,另使用MTT法检测细胞活性。结果 成功建立的MDM细胞模型对Beads和细菌的吞噬呈时效关。Rolipram在实验浓度(10^~8~10^-5 M)下对MDM吞噬Beads、H.influenzae和S.aureus能力无明显促进或抑制作用,也不影响MDM的活性。结论 磷酸二酯酶4抑制剂不会因升高巨噬细胞内cAMP水平而影响其吞噬非生物颗粒和细菌的能力。

Objective To investigate the influence of phosphodiesterases 4 inhibitor on the phagocytosis of non-biological particles and gram-positive bacteria, gram-negative bacteria by human alveolar macrophages. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from venous blood from 12 healthy volunteers using Ficoll-Hypaque density gradients. Monocytes were incubated with media containing 2 ng/ml GM-CSF for 12d to allow full differentiation into macrophage (MDM), a functionally equivalent model of human AM. MDM were pretreated with Rolipram overnight (16-18h),phagocytosis of fluorescent labeled beads and H.influenzae,S.aureus by MDM was measured using a fluostar optima fluorimeter. Cell viability was assay with MTT. Results MDM phagocytosis of beads and bacteria was a time-dependant process. Rolipram in the concentration of 10^-8-10^-5M didn't inhibit or promote phagocytosis of beads and bacteria by MDM, and didn't affect the cell viability. Conclusion Phosphodiesterases 4 inhibitor would not affect the human macrophage phagocytic capacity of non-biological particles and bacteria associated with enhanced intracellular cAMP level.

目的 探讨GDM孕早、中期脂肪细胞因子Chemerin、RBP4水平的变化。方法 采用前瞻性研究,测定38例GDM及40例正常孕妇孕早、中期血清Chemerin、RBP4水平,分析其与胰岛素抵抗的相关性。结果 GDM组孕妇孕早、中期血清Chemerin、RBP4水平及HOMA-IR均较对照组升高,差异有统计学意义(P<0.05);两组孕妇孕中期血清Chemerin、RBP4水平均较孕早期升高,差异有统计学意义 (P<0.05);Chemerin、RBP4水平与IR成正相关,差异有统计学意义(P<0.05)。结论 GDM孕妇孕早、中期脂肪细胞因子Chemerin、RBP4水平升高,Chemerin、RBP4水平的升高与IR有一定相关性。

Objective To discuss the serum levels of the Adipokine Chemerin、RBP4 of Gestational Diabetes Mellitus(GDM) in early pregnancy and middle pregnancy. Methods In prospective study, pregnant women, venous blood was collected from 40 controls and 38 GDM during early pregnancy (9-12weeks)and middle pregnancy (22-26weeks). Serum insulin, Chemerin, RBP4 were measured by enzyme-linked immunosorbent assay (ELISA). Results Mean serum levelsof Chemerin and RBP4 were significantly higher among GDM cases compared with controls during early pregnancy and middle pregnancy(P<0.05). In two groups, Mean serum levels of Chemerin and RBP4 in middle pregnancy were significantly higher than those in early pregnancy (P<0.05).During early pregnancy and middle pregnancy, the Chemerin and RBP4 levels were positively related with HOMA-IR (P＜0.05). Conclusion There is evidence of a positive association elevated Chemerin and RBP4 concentration of early pregnancy and middle pregnancy with increased GDM risk.

目的 构建抑癌基因SEMA3B真核表达载体pcDNA3.1-SEMA3B,并检测其对肺癌A549细胞恶性生物学行为的影响。方法 应用PCR扩增SEMA3B全长cDNA片段,构建真核表达载体pcDNA3.1-SEMA3B。克隆PCR、双酶切法、基因测序验证过表达载体构建成功。将pcDNA3.1-SEMA3B真核表达载体和空载体pcDNA3.1分别转染入A549细胞中,应用qRT-PCR、Western blot检测SEMA3B mRNA、蛋白表达水平的变化;MTS法检测细胞增殖;流式细胞仪检测细胞凋亡、细胞周期;克隆形成实验检测细胞集落形成能力。结果 SEMA3B基因扩增片段与预测片段一致,克隆成功,且测序鉴定证实真核表达载体构建成功。转染pcDNA3.1-SEMA3B真核表达载体可上调SEMA3B mRNA、蛋白表达水平,且可抑制A549细胞的增殖,诱导凋细胞亡,细胞被阻滞在G1期,抑制细胞集落形成能力。结论 成功构建了SEMA3B基因真核表达载体,抑癌基因SEMA3B在肺癌恶性生物学进程中可能发挥重要作用。

Objective To construct the eukaryotic expression vector of the cancer suppressor gene, SEMA3B, and research the effects on malignant biological behavior of lung cancer A549 cells. Methods By reverse transcriptase-polymerase chain reaction (RT-PCR), the full length SEMA3B gene was amplified and then was inserted into pcDNA3.1. The recombinant plasmid pcDNA3.1-SEMA3B was confirmed correctly through double enzyme digestion and PCR identification, which was transfected into lung cancer A549 cells by lipid media transfection. The untransfected A549 and A549 transfected with pcDNA3.1 were used as controls. SMEA3B gene was detected by qRT-PCR and western blot. MTS assay, flow cytometry, and colony formation test were performed to evaluate the effect of overexpression of SEMA3B gene on A549 cell proliferation, apoptosis, cell cycle, and colony forming ability. Results The amplied fragment of SEMA3B gene by PCR was consistent with the anticipated result, the SEMA3B gene was cloned successfully. And the recombinant plasmid pcDNA3.1-SMEA3B was constructed successfully through gene sequence identification. After transfection of pcDNA3.1-SEMA3B, SEMA3B mRNA and protein expression levels were raised, and overexpression of SEMA3B gene in A549 cells significantly inhibited the proliferation of A549 cells, induced apoptotic cell death, blocked cell cycle in the G1 phase, and suppressed cell colony-forming ability. Conclusion The recombinant pcDNA3.1-SEMA3B is constructed successfully. SEMA3B gene can significantly inhibit the malignant biological behavior of lung cancer A549 cells.

目的 分析乳腺癌细胞中Snail与MTDH基因的作用,明确Snail是否通过结合于MTDH的启动子区域促进乳腺癌转移。方法 克隆、转染Snail基因至乳腺癌细胞,观察过表达Snail的乳腺癌细胞中MTDHmRNA及蛋白表达的变化;再使用免疫共沉淀法检测Snail与MTDH基因的共作用。结果 转染Snail基因进入乳腺癌MDA-MB-435细胞后,转染组、空白组和对照组中MTDHmRNA的表达水平分别为1.61±0.22、1.02±0.18、0.99±0.20,转染组高于空白组和对照组,差异有统计学意义(P<0.05),而后两组表达无差异(P>0.05);Westren blot检测结果显示,Snail可促进MTDH蛋白的表达;免疫共沉淀显示,Snail与MTDH在细胞内存在相互结合作用。结论 Snail在乳腺癌细胞中可通过结合于MTDH基因的启动子区域,促进MTDHmRNA转录及相关蛋白的表达,从而导致乳腺癌转移。

Objective To investigate the function of Snail to MTDH gene in breast cancer cells. Methods We observed the changement of MTDHmRNA and protein expression in breast cancer cell line MDA-MB-435 after transfected with Snail gene. Then, we used co-immunoprecipitation to determine the domain of Snail and MTDH binding in vitro. Results After transfected with Snail gene into MDA-MB-435 cell, the expression levels of MTDHmRNA in transfection group, blank group and control group were 1.61±0.22,1.02±0.18,0.99±0.20. The level of transfection group was significantly higher than the other groups(P< 0.05). Western blot showed that the expression of MTDH protein can be promoted by Snail. Co-immunoprecipitation showed that Snail and MTDH are binding interactions in breast cancer cell line MDA-MB-43. Conclusion Snail can promote transcription and expression of MTDH in breast cancer cells by binding to the promoter region of the MTDH gene resulting in metastasis of breast cancer.

目的 建立大鼠急性心肌梗死缺血再灌注后无复流模型,并初步验证细胞焦亡在其中的发生情况。方法 选用20只标准成年雄性Sprague Dawley大鼠（体质量260~320 g）,随机分为对照组（n=5）和手术组（n=15）。对照组仅穿线冠状动脉,未行结扎;手术组结扎左前降支0.5 h后解除,进行再灌注4 h,以建立无复流模型。通过Evens blue和硫磺素S染色,评估心肌的正常供血区、再灌注区及无复流区,并对两组大鼠心肌组织进行病理分析。结果 对照组大鼠全部存活,未出现无复流现象,心肌组织中未见细胞焦亡。手术组存活13只,形成明确的正常供血区（n=13）、再灌注区（n=13）和无复流区（n=10）。在无复流区的心肌细胞中均观察到细胞焦亡（n=10）,而正常供血区未见（n=0）,再灌注区部分出现（n=4）,差异具有统计学意义（P<0.05）。结论 细胞焦亡现象主要存在于大鼠急性心肌梗死缺血再灌注后无复流区域中,细胞焦亡可能作为一种区域特异性程序性死亡方式,在心肌无复流的发生与发展中发挥重要作用。

Objective To establish a rat model of myocardial no-reflow after acute myocardial infarction with ischemia-reperfusion injury and to preliminarily explore the occurrence of pyroptosis in the affected myocardium. Methods Twenty adult male Sprague-Dawley rats（260-320 g）were randomly divided into a control group（n=5）and a surgical group（n=15）. In the control group,the coronary artery was encircled with suture but not ligated. In the surgical group,the left anterior descending artery was ligated for 30 minutes, followed by 4 hours of reperfusion to induce the no-reflow model. Evans blue and thioflavin S staining were used to evaluate the normal perfusion area,reperfusion area,and no-reflow area of the myocardium. Histopathological analysis was conducted on myocardial tissues from both groups. Results All rats in the control group survived without evidence of no-reflow or pyroptosis in myocardial tissue. In the surgical group, 13 rats survived and showed distinct regions of normal perfusion, 13 with reperfusion, and 10 with no-reflow. Pyroptosis was observed in all no-reflow areas（n=10）, absent in the normal perfusion zones（n=0）, and partially present in the reperfusion zones（n=4）. The differences were statistically significant（P<0. 05）. Conclusions Pyroptosis predominantly occurs in the no-reflow zones following acute myocardial infarction and ischemia-reperfusion injury in rats. As a region-specific form of programmed cell death, pyroptosis may play an important role in the development of myocardial no-reflow.

目的 初步探究胆管结扎诱导的梗阻性胆汁淤积对大鼠肝细胞的影响。方法 10只Lewis大鼠随机分为对照组和胆汁淤积组,每组各5只,胆汁淤积组采用胆管结扎2周诱导梗阻性胆汁淤积大鼠模型。苏木精-伊红染色和苯胺蓝染色比较组织病理变化,使用生化分析比较两组小鼠肝功能情况。采用改良的两步胶原酶灌注分离原代肝细胞,通过RT-qPCR检测两组小鼠肝细胞标志基因、细胞增殖标志基因以及胆管细胞标志基因的表达情况。结果 与对照组相比,胆汁淤积组肝脏表现为明显的肝组织紊乱和纤维胶原蛋白沉积以及肝功能的损害。胆汁淤积组较对照组的原代肝细胞更高表达细胞增殖标志基因：细胞增殖标志物（Ki67）基因,叉头盒M1蛋白（Foxm1）基因,增殖细胞核抗原（Pcna）基因和肝细胞生长因子（HGF）基因（P<0.05）;胆汁淤积组的原代肝细胞表达更低水平的肝细胞标志基因：白蛋白（Alb）基因,多药耐药相关蛋白2（Mrp2）基因,胆盐输出泵（Bsep）基因和肝细胞连环蛋白1（Catenin1）基因（P<0.05）,同时表达更高水平的胆管细胞标志基因：细胞角蛋白7（Ck7）基因,细胞角蛋白 19（Ck19）基因,胆管细胞多药耐药性蛋白1（Mdr1）基因和胆管细胞囊性纤维化跨膜传导调节因子（Cftr）基因（P<0.05）以及肝祖细胞标志基因：上皮细胞黏附分子（Epcam）基因和Y染色体性别决定区-盒转录因子9（Sox9）基因（P<0.05）。结论 胆汁淤积可诱导肝细胞向胆管细胞特性转化的可塑性。

Objective To explore the effect of bile duct ligation induced obstructive cholestasis on rat hepatocytes. Methods Ten Lewis rats were randomly divided into control group and cholestasis group, and the cholestasis was induced by bile duct ligation for 2 weeks. The histopathological changes were compared by H&E and aniline blue staining and the liver function was compared by biochemical analysis. Primary hepatocytes were isolated by modified two-step collagenase perfusion, and the expressions of hepatocyte marker genes, cell proliferation marker genes and cholangiocyte marker genes were detected by RT-qPCR. Results Compared with the control group,the liver of the cholestatic group showed obvious disordered histopathology, deposition of fibrous collagen and impaired liver function. Compared with the control group, the primary hepatocytes in the cholestasis group expressed higher cell proliferation-related genes（Ki67,Foxm1,Pcna and HGF）（P<0. 05）. Primary hepatocytes in the cholestasis group expressed lower levels of hepatocyte marker genes（Alb,Mrp2,Bsep and Catenin1）（P<0. 05）,and higher levels of cholangiocyte marker genes（Ck7,Ck19,Mdr1 and Cftr）（P<0. 05）and higher levels of the hepatic progenitor cell marker genes（Epcam and Sox9）（P<0. 05）. Conclusions Cholestasis induces rat hepatocyte plasticity in the transformation into bile duct properties.

       目的   评估无托槽隐形矫治应用在正畸拔牙患者中的效果及对牙根吸收、可溶性细胞间黏附分子-1（sICAM-1）的影响。方法   纳入2022年1月—2024年8月的70例正畸拔牙患者，按照治疗方法分组，即对照组（35例，给予固定矫治）、观察组（35例，给予无托槽隐形矫治），评价组间牙根吸收情况、牙周指标、炎症因子、矫治时间。结果   治疗结束时，两组均出现牙根吸收情况，但是观察组无牙根吸收＞3 mm病例，而对照组存在牙根吸收＞3 mm、＞4 mm病例，P＜0.05。治疗前，两组牙周指标［龈沟出血指数（SBI）、牙龈指数（GI）、菌斑指数（PLI）］、炎症因子［白介素-1β（IL-1β）、sICAM-1］比较差异无统计学意义（P＞0.05）。治疗后，两组SBI、GI、PLI、IL-1β、sICAM-1升高，且观察组SBI、GI、PLI、IL-1β、sICAM-1低于对照组（P＜0.05）。与对照组比较，观察组矫治时间更长（P＜0.05）。结论   对正畸拔牙患者进行无托槽隐形矫治，虽然治疗时间长，但是可以抑制牙根吸收，减轻炎症反应，提高牙周健康水平。

       Objective  To evaluate the effect of clear aligner treatment on orthodontic tooth extraction patients and its impact on root resorption and soluble intercellular adhesion molecule-1（sICAM-1）．Methods  Seventy orthodontic extraction patients from January 2022 to August 2024 were included and divided into two groups according to treatment methods：a control group（35 cases，receiving fixed orthodontic treatment）and an observation group（35 cases，receiving clear aligner treatment）．The root resorption，periodontal indicators，inflammatory factors，and orthodontic treatment time between groups were evaluated．Results  At the end of treatment，both groups showed root resorption，but there were no cases of root resorption＞3 mm in the observation group，while there were cases of root resorption＞3 mm and＞4 mm in the control group，P＜0.05．Before treatment，there was no difference in periodontal indicators（gingival bleeding index［SBI］，gingival index［GI］，plaque index［PLI］），inflammatory factors（interleukin-1 β［IL-1 β］，sICAM-1） between the groups，P＞0.05．After treatment，SBI，GI，PLI，IL-1 β，sICAM-1 increased in both groups，but SBI，GI，PLI，IL-1 β，sICAM-1 were lower in the  observation group，P＜0.05．Compared with the control group，the observation group had a longer orthodontic treatment time，P＜0.05．Conclusions  Although the clear aligner treatment time for orthodontic extraction patients is longer，it can inhibit root resorption，reduce inflammatory reactions，and improve periodontal health．