大鼠Caspase 8基因启动子荧光素酶报告质粒的构建及其与IRF-1结合位点的鉴定

目的:构建大鼠Caspase 8基因启动子(全长和截短)荧光素酶报告质粒,并观察在人胚肾细胞(HEK293)中,过表达干扰素调节因子-1(interferon regulatory factor-1,IRF-1)对Caspase 8基因启动活性的影响?同时,筛选其可能的IRF-1结合位点?方法:采用PCR技术,扩增出大鼠Caspase 8基因启动子序列(-1136~+101 nt),将Caspase 8基因启动子插入到荧光素酶报告基因载体pGL3-basic中?将Caspase 8基因启动子全长荧光素酶报告质粒(pGL3-Caspase 8-FL)和大鼠野生型IRF-1表达质粒(pcDNA3.1-IRF-1)共转染HEK293细胞,检测其荧光素酶活性,确定IRF-1对Caspase 8基因的启动作用?另用生物信息学软件预测Caspase 8基因启动子上IRF-1潜在的结合位点,并构建Caspase 8基因启动子截短的荧光素酶报告质粒(即pGL3-Caspase 8-1~4)?将上述Caspase 8基因启动子全长和各截短的荧光素酶报告质粒和IRF-1过表达质粒共转染HEK293细胞,再行荧光素酶活性测定,筛选IRF-1的结合位点?结果:菌液PCR及核酸测序证实,上述荧光素酶报告质粒均构建成功?将pGL3-Caspase 8-FL和pcDNA3.1-IRF-1共转染HEK293细胞发现,Caspase 8基因启动子活性显著增加?而将pGL3-Caspase 8-FL?pGL3-Caspase 8-1~4和pcDNA3.1-IRF-1共转染HEK293细胞后证实,pGL3-Caspase 8-4的启动活性显著低于pGL3-Caspase 8-2和pGL3-Caspase 8-3?提示IRF-1可能结合在Caspase 8基因启动子的-336~-136 nt区域?结论:本实验成功构建了大鼠Caspase 8基因启动子全长及截短荧光素酶报告质粒,并初步筛查出IRF-1在Caspase 8基因启动子上的结合区域?     

TLR7刺激对系统性红斑狼疮易感型小鼠树突状细胞免疫功能的影响

目的 研究TLR7通路对于系统性红斑狼疮易感Fas - -M RLIpr/Ipr小鼠树突状细胞免疫功能的影响.方法 从5周龄MRLIpr/Ipr小鼠骨髓提取DC体外培养,以TLR7激活剂咪喹莫特处理48h,流式细胞仪检测细胞表面分子CD80、MHCⅡ表达.ELISA法检测细胞培养上清中细胞因子IFN-γ、TNF-α、IL-10.结果 咪喹莫特处理后,树突状细胞表面抗原提呈相关分子MHCⅡ和CD80分子均降低,分泌的IFN-γ、TNF-α和IL-10均明显增加.结论TLR7通路调节系统性红斑狼疮易感小鼠MRLIpr/Ipr小鼠DC抗原呈递能力,具有影响免疫应答作用.     

抗精子抗体ELISA检测法及其应用

建立了用精子可溶性抗原包被的ELISA法,检测了100例处女血清,均为阴性,17例有生育能力者和1000例不育患者血清的阳性率分别为5.9％和22％,抗体检出率符合国外多数学者的结果。重复性试验定性符合率达97％。表明用该法检测抗精子抗体具有敏感、特异、稳定等特点,可获较为满意的结果。     

小儿脑瘫的推拿治疗

目的:观察推拿手法治疗小儿脑瘫的疗效.方法:选取背腰部穴区、头颈部穴区、四肢部穴区进行手法治疗;益气健脑、理筋整复.及早发现,及早诊断、及早治疗,达最佳治疗效果.结果:推拿手法可益气通经,健脑补肾,改善头部血液循环,促进血栓吸收,达到治病效果.结论:推拿手法治疗小儿脑瘫可以起到较好的效果,值得推广.     

米利酮对麻醉家兔的心血管作用

静脉注射米利酮(3～300μg/kg),能使麻醉家兔正常心脏左室内压最大上升和下降速率(LV士dP/dt max)呈剂量依赖性增加,心率(HR)仅在高剂量时略有增加,而全身动脉压(AP)无明显变化。米利酮在静脉剂量300μg/kg才增加呼吸振幅和频率。给家兔灌胃米利酮(10mg/kg,2次/d)一周后,其心脏收缩和舒张功能均显著高于对照组。米利酮(500μg/kg iv)可明显翻转家兔戊巴比妥钠诱导的衰竭心脏血流动力学指标,AP亦无变化。结果表明,米利酮对家兔正常和衰竭心脏都有明显的正性肌力作用,且在这些剂量不改变心脏后负荷。     

微创交锁髓内钉技术治疗股骨干骨折的疗效分析

目的:探讨全闭合复位交锁髓内钉固定,配合股骨远端徒手锁钉技术治疗股骨干骨折的疗效。方法:采用全闭合复位髓内钉固定治疗股骨骨折,辅以股骨远端徒手锁钉技术治疗13例股骨干骨折患者,共18例股骨,男9例,女4例;左侧10例,右侧8例;年龄17-53岁,平均36.3岁。根据AO分型:A1型3例,A2型3例,A3型2例;B1型2例,B2型5例,B3型1例;C1型1例,C2型1例。记录手术时间、术中出血量、术中透视时间、术后近期并发症及术后住院时间等。结果:手术时间平均为60(45-135)min,出血量平均为155(50-280)ml。术中透视时间平均为95(50-170)s。无中转为切开复位内固定病例,平均住院时间为15(14-18)d。1例双股骨干骨折患者术后并发脂肪栓塞,治疗后痊愈。所有患者术后获6-18个月(平均10个月)随访,骨折临床愈合时间为5-11个月(平均8个月)。对膝关节功能评定按照Klemm功能恢复标准评价术后疗效,优14例,良4例,优良率100%。无明显的术后近期并发症。结论:全闭合复位徒手锁钉技术治疗股骨干骨折具有手术时间短、出血少、创伤小,患肢功能及骨折愈合快等优点,进一步发挥了交锁髓内钉优势,效果良好。     

恒速滴注法在高龄卧床管饲患者中的应用效果

目的探讨恒速滴注法在高龄卧床管饲患者中的应用效果。方法2010年5月至2011年8月，方便性抽样选择在解放军总医院住院治疗的携带鼻胃管或鼻肠管行胃肠营养的老年患者60例，按入院先后分为观察组和对照组各30例，观察组采用恒速滴注法管饲肠内营养乳剂，对照组采用常规管饲方法即分次推注法，观察并比较两组患者堵管、腹胀、腹泻、反流、误吸等并发症的发生率。结果观察组患者发生堵管、反流及误吸的例数均少于对照组，差异均有统计学意义（均P〈0．05）；发生腹胀与腹泻的例数与对照组比较，差异无统计学意义（均P〉0．05）。结论恒速滴注法肠内营养可有效满足高龄长期卧床患者的营养需求，预防和减少并发症的发生，推荐在临床广泛应用。     

基于DEA模型的生活垃圾转运站产能及效率评价的初步研究

结合最新颁布实施的CJJ/T 156—2010生活垃圾转运站评价标准,选取相应评价指标,采用DEA模型对垃圾转运站的产能及效率进行评价,并通过MATLAB程序,使评价模型操作简单化。     

Comparison of the Masaoka-Koga staging and the International Association for the Study of Lung Cancer/the International Thymic Malignancies Interest Group proposal for the TNM staging systems based on the Chinese Alliance for Research in Thymomas retrospective database

Background

To compare the predictive effect of the Masaoka-Koga staging system and the International Association for the Study of Lung Cancer (IASLC)/the International Thymic Malignancies Interest Group (ITMIG) proposal for the new TNM staging on prognosis of thymic malignancies using the Chinese Alliance for Research in Thymomas (ChART) retrospective database.

Methods

From 1992 to 2012, 2,370 patients in ChART database were retrospectively reviewed. Of these, 1,198 patients with complete information on TNM stage, Masaoka-Koga stage, and survival were used for analysis. Cumulative incidence of recurrence (CIR) was assessed in R0 patients. Overall survival (OS) was evaluated both in an R0 resected cohort, as well as in all patients (any R status). CIR and OS were first analyzed according to the Masaoka-Koga staging system. Then, they were compared using the new TNM staging proposal.

Results

Based on Masaoka-Koga staging system, significant difference was detected in CIR among all stages. However, no survival difference was revealed between stage I and II, or between stage II and III. Stage IV carried the highest risk of recurrence and worst survival. According to the new TNM staging proposal, CIR in T1a was significantly lower comparing to all other T categories (P<0.05) and there is a significant difference in OS between T1a and T1b (P=0.004). T4 had the worst OS comparing to all other T categories. CIR and OS were significantly worse in N (+) than in N0 patients. Significant difference in CIR and OS was detected between M0 and M1b, but not between M0 and M1a. OS was almost always statistically different when comparison was made between stages I–IIIa and stages IIIb–IVb. However, no statistical difference could be detected among stages IIIb to IVb.

Conclusions

Compared with Masaoka-Koga staging, the IASLC/ITMIG TNM staging proposal not only describes the extent of tumor invasion but also provides information on lymphatic involvement and tumor dissemination. Further study using prospectively recorded information on the proposed TNM categories would be helpful to better grouping thymic tumors for predicting prognosis and guiding clinical management.     

The prognostic value of inflammation markers in postoperative gliomas with or without adjuvant treatments

Recent studies have shown that some inflammatory markers are associated with the prognosis of solid tumors. This study aims to evaluate the prognosis of glioma patients with or without adjuvant treatment using the systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), and platelet-lymphocyte ratio (PLR).All patients who were diagnosed with gliomas at the first and second affiliated hospital of Guangxi Medical University between 2011 and 2020 were included in this study. The optimal cutoff value of SII, NLR, and PLR was determined by X-tile software program. We stratified patients into several groups and evaluated the progression-free survival (PFS) and overall survival (OS) of SII, NLR, and PLR during the period of pre-surgical, con-chemoradiotherapy, and post-treatments. Multivariate Cox regression analyses were performed to detect the relationships between OS, PFS, and prognostic variables.A total of 67 gliomas patients were enrolled in the study. The cutoff values of SII, NLR, and PLR were 781.5 × 10⁹/L, 2.9 × 10⁹/L, and 123.2 × 10⁹/L, respectively. Patients who are pre-SII < 781.5 × 10⁹/L had better PFS (P = .027), but no difference in OS. In addition, patients who had low pre-NLR (<2.9 × 10⁹/L) meant better OS and PFS. PLR after adjuvant treatments (post-PLR) was significantly higher than pre-PLR (P = .035). Multivariate analyses revealed that pre-SII, pre-NLR were independent prognostic factors for OS (pre-SII: HR 1.002, 95% CI: 1.000–1.005, P = .030 and pre-PLR: HR 0.983, 95% CI: 0.973–0.994, P = .001), while pre-PLR was an independent factor for PFS (HR 0.989, 95% CI: 0.979–1.000, P = .041).High pre-SII or high pre-NLR could be prognostic markers to identify glioma patients who had a poor prognosis.