Changes of mesenchymal stromal cells mobilization and bone turnover in an experimental bone fracture model in ovariectomized mice

Objective: The aim of this study was to characterize the mesenchymal stromal cells (MSCs) and endothelial progenitor cells (EPCs) mobilization, and bone turnover in osteoporotic fracture healing in ovariectomized mice. Methods: In total, 112 female C57/BL mice were divided into two groups. The first group was sham-operated (SO), and the other group was ovariectomized (OVX). After three weeks, the right femora of the mice were fractured under anesthesia and internally fixed with steel pin. Peripheral blood and bone marrow were was collected for flow cytometry analysis, at 0 hours (h), 12 h, 24 h, 72 h and 168 h after fracture. MSCs and EPCs levels were assessed using cell surface antigens in different combinations (CD44+ CD34-CD45-, and CD34+ KDR+CD45-) by flow cytometry. At 0, 14, 28 and 42 days after fracture, sera were assayed for circulating levels of procollagen type I-N-terminal propeptide (P1NP) and C-terminal telopeptide of type I-collagen (CTX) by ELISA. Femurs were harvested at 2 weeks and 6 weeks after fracture for X-ray radiography, micro-computed tomography (micro-CT) and histology. Results: Our results showed that bone marrow and peripheral blood MSCs numbers of the OVX mice were significantly lower than the SO mice, at 12 h, 24 h and 72 h after fracture. In addition, circulating P1NP and CTX levels of the OVX mice were significantly higher than the SO mice, at 2 and 4 weeks. Conclusion: Results of the present study revealed disorders of bone marrow MSCs mobilization and bone turnover may partially account for the delay of osteoporotic fracture healing.     

Rare presentation of multi-organ abdominal echinococcosis: report of a case and review of literature

Hydatid disease, which is also known as cystic echinococcosis, is a zoonotic infection caused by the cestode tapeworm Echinococcus granulosus and rarely by Echinococcus multilocularis. In this report we describe an unusual case of a 19-year-old woman who was admitted to our hospital for abdominal pain, nausea, and vomiting. Computed tomography revealed multi-organ abdominal echinococcosis. The patient recovered after undergoing surgery to excise the cyst. The diagnosis, clinical features, treatment, and prevention in this case of multi-organ abdominal echinococcosis are discussed, in light of the relevant literature.     

Significance of Fas and FasL protein expression in cardiac carcinoma and local lymph node tissues

Objective: To investigate the relation of Fas and Fas ligand (FasL) protein expression with carcinogenesis and metastasis of cardiac carcinoma. Methods: Immunohistochemistry was used to detect Fas and FasL protein expression in 64 cardiac carcinoma tissue samples and 20 normal gastric tissue samples. Relation between FasL and Fas expression, age and gender of gastric cancer patients, and pathological subtype and lymph node metastasis of gastric cancer was analyzed. Results: The Fas expression level was significantly higher in normal gastric tissue samples than in cardiac carcinoma tissue samples (85.0% vs. 25.0%, P<0.001), while the FasL expression level was significantly lower in normal gastric tissue samples than in cardiac carcinoma tissue samples (30.0% vs. 81.3%, P<0.001). The Fas expression level was significantly higher in invasive lymph nodes than in non-invasive lymph nodes (82.9% vs. 56.5%, P<0.003) and in well-differentiated gastric carcinoma tissue samples than in poorly-differentiated cardiac carcinoma tissue samples (50.0% vs. 18.0%, P=0.015). The FasL expression level was significantly lower in well-differentiated cardiac carcinoma tissue samples than in poorly- differentiated cardiac carcinoma tissue samples (42.9% vs. 84.0%, P=0.021). The Fas and FasL expression levels (25.0% and 81.3%) were significantly different in cardiac carcinoma tissue samples (P<0.001), but had a non-linear correlation (P=0.575). Conclusion: Abnormal Fas and FasL expressions in cardiac carcinoma and lymph node tissues are involved in carcinogenesis and metastasis of gastric cancer.     

Cross-Protection of Influenza A Virus Infection by a DNA Aptamer Targeting the PA Endonuclease Domain

Amino acid residues in the N-terminal of the PA subunit (PA_N) of the influenza A virus polymerase play critical roles in endonuclease activity, protein stability, and viral RNA (vRNA) promoter binding. In addition, PA_N is highly conserved among different subtypes of influenza virus, which suggests PA_N to be a desired target in the development of anti-influenza agents. We selected DNA aptamers targeting the intact PA protein or the PA_N domain of an H5N1 virus strain using systematic evolution of ligands by exponential enrichment (SELEX). The binding affinities of selected aptamers were measured, followed by an evaluation of in vitro endonuclease inhibitory activity. Next, the antiviral effects of enriched aptamers against influenza A virus infections were examined. A total of three aptamers targeting PA and six aptamers targeting PA_N were selected. Our data demonstrated that all three PA-selected aptamers neither inhibited endonuclease activity nor exhibited antiviral efficacy, whereas four of the six PA_N-selected aptamers inhibited both endonuclease activity and H5N1 virus infection. Among the four effective aptamers, one exhibited cross-protection against infections of H1N1, H5N1, H7N7, and H7N9 influenza viruses, with a 50% inhibitory concentration (IC₅₀) of around 10 nM. Notably, this aptamer was identified at the 5th round but disappeared after the 10th round of selection, suggesting that the identification and evaluation of aptamers at early rounds of selection may be highly helpful for screening effective aptamers. Overall, our study provides novel insights for screening and developing effective aptamers for use as anti-influenza drugs.     

Do Formulation Differences between the Reference Listed Drug and Generic Piperacillin-Tazobactam Impact Reconstitution?

Pharmaceutical differences between the reference listed drug (RLD) and generic formulations of piperacillin-tazobactam may impact the reconstitution process for intravenous administration. This study evaluated the RLD against three generic formulations and measured their reconstitution times using a standardized process. The mean (standard deviation [SD]) reconstitution time for one generic formulation was 5.57 (1.49) min, which was 35% to 42% longer (P < 0.002) than that for the RLD and two other formulations. Observable microscopic differences in powder particle morphology may explain these findings.     

应用siRNA敲低TPM3基因表达后对大鼠胰腺癌细胞侵袭转移能力的影响

目的：探究原肌球蛋白3（tropomyosin alpha-3 chain,TPM3）在大鼠胰腺癌细胞中对其侵袭转移能力的影响并初步阐述其可能的分子机制。方法：（1）将70只大鼠随机分为手术组40只[将7,12-二甲基苯并蒽（7,12-dimethyl-1,2-benzanthracene,DMBA]植入大鼠胰腺被膜下后荷包缝合被膜）,假手术组15只（只对胰腺进行荷包缝合而后关腹,不置入DMBA）,阴性对照组15只（不做任何处理）,建立大鼠胰腺癌模型。（2）通过组织免疫组化染色的方法对差异蛋白TPM3进行验证。（3）通过机械分离和酶阶段消化法分离胰腺癌组织的方法获取大鼠胰腺癌细胞,体外传代培养获得纯度较高的细胞后,应用siRNA敲低大鼠胰腺癌细胞中TPM3基因的表达;通过RT-PCR技术检测其沉默效果。（4）通过Transwell实验和平板克隆实验分别对细胞的侵袭、迁移能力及生长增殖能力进行观察。结果：（1）经病理验证模型组有37.83%（14/37）形成胰腺癌,证明大鼠胰腺癌模型建立成功。（2）免疫组化验证胰腺癌组织中TPM3阳性率（92.8%）明显高于正常胰腺组织（6.7%）。（3）RT-PCR结果显示,转染TPM3-siRNA的细胞中TPM3的表达量（0.31±0.02）明显低于其余各组,脂质体组（0.45±0.02）,阴性对照组（0.45±0.02）,空白对照组（0.43±0.02）（P＜0.05）,其余各组之间TPM3表达量差异无统计学意义（P ＞0.05）。（4）转染TPM3-siRNA的细胞其侵袭[（161.63±4.94）个]、迁移能力[（206.87±4.21）个]及生长增殖能力[（51.5±2.327）个]较对照组明显降低[分别为（39.7±1.40）个,（67.27±1.76）个,（5.900±0.767）个]（P＜0.05）。结论：体外化学合成的TPM3-siRNA可有效的抑制大鼠胰腺细胞TPM3的表达,TPM3表达降低后其侵袭能力、迁移能力及生长增殖能力下降。     

人肝再生增强因子真核表达及其对顺铂诱导肝癌细胞凋亡的影响

目的：构建人肝再生增强因子（human augmenter of liver regeneration,hALR）15kD亚型的真核表达系统,并探索ALR对顺铂（Cisplatin,DDP）诱导的人肝癌细胞株QGY凋亡的影响。方法：用聚合酶链反应（polymerase chain reaction,PCR）方法和基因重组技术,从重组质粒pPIC9K-rhALR中扩增出编码rhALR基因片段,将其克隆入pPICZαA质粒中构建重组质粒pPICZα－A-rhALR。重组质粒SacⅠ酶切线性化后电转入酵母GS115中,并在1%的甲醇诱导下表达。表达上清蛋白经Western blot（ALR多克隆抗体和His-tag标签抗体）鉴定和镍柱亲和层析纯化。MTS试剂检测rhALR体外对人肝癌细胞（HepG2、QGY）的促增殖活性,及流式细胞仪检测rhALR在顺铂诱导QGY细胞凋亡中的抗凋亡作用。结果：PCR、双酶切、DNA测序均鉴定重组质粒构建与预期一致。分泌表达的rhALR约占上清总蛋白的70%,目的分子量约17 kD,Western blot均可见单一条带。纯化后的rhALR对HepG2和QGY的体外促增殖作用呈浓度依赖性增强（HepG2组：F=246.729,P=0.000;QGY组：F=246.004,P=0.000）,且在QGY细胞顺铂诱导凋亡时也发挥着浓度梯度式抗凋亡作用（F=101.061,P=0.000）。结论：成功构建高效分泌表达rhALR的pPICZα－A-rhALR GS115真核表达系统;体外实验证实rhALR对顺铂诱导人肝癌细胞有呈浓度依赖性的抗凋亡作用。     

第三脑室颅咽管瘤的手术治疗策略

目的:总结第三脑室颅咽管瘤的手术治疗策略。方法:回顾性分析行手术治疗的28例第三脑室颅咽管瘤患者的临床资料,对比经终板和经前纵裂-胼胝体-透明隔-穹窿间2种手术入路的治疗效果和适应症。结果:2种手术入路在全切肿瘤率上无明显的统计学差异(P=0.907),经前纵裂-胼胝体-透明隔-穹窿间入路比经终板入路更容易出现尿崩(P=0.027)、电解质紊乱(P=0.033)等并发症。经终板入路适用肿瘤主体位于视交叉后上第三脑室底前部的肿瘤切除,而经前纵裂-胼胝体-透明隔-穹窿间入路适用于完全位于第三脑室内或肿瘤主体位于第三脑室内的肿瘤切除。结论:根据颅咽管瘤的影像学特征选择适当的手术入路,术中注意保护下丘脑和垂体柄是手术成功的关键。     

肱骨近端锁定加压钛板治疗老年肱骨外科颈骨折的疗效观察

目的：探讨应用肱骨近端锁定加压钛板系统治疗老年肱骨外科颈骨折的疗效。方法总结2007年8月至2012年8月采用肱骨近端锁定加压钛板系统切开复位治疗老年肱骨外科颈骨折32例的临床资料,男13例,女19例;年龄59～78岁,平均66.7岁;Neer分型,二部分骨折7例,三部分骨折16例,四部分骨折9例。结果术后X线显示钛板螺钉位置良好,骨折复位满意,无神经血管损伤。手术时间平均（58±24）min,术中出血量平均（135±28）ml。根据Neer肩关节功能评分系统进行评分,优21例,良8例,可2例,差1例,优良率达90.6％。结论肱骨近端锁定加压钛板具有置钉角度科学、固定可靠、可早期行功能锻炼等优点,较适用于老年骨质疏松粉碎性肱骨外科颈骨折的治疗。     

241例重庆地区急性呼吸道感染患儿肺炎链球菌耐药性分析

目的:了解重庆地区儿童急性呼吸道感染肺炎链球菌(Streptococcus pneumoniae,SP)的耐药情况。方法:采集重庆医科大学附属儿童医院2014年1~3月、9~11月急性呼吸道感染患儿呼吸道标本(痰、支气管肺泡灌洗液),分离培养,得到SP进行药敏检测。结果:共收集241株SP,红霉素耐药率最高,达97.10%,四环素91.70%、克林霉素89.63%,未检出万古霉素耐药株;利奈唑胺、氯霉素、泰利霉素、左旋氧氟沙星、莫西沙星敏感率较高,达90%以上;青霉素、美洛培南、头孢吡肟的不敏感率分别为47.3%、81.74%、67.22%;主要耐药模式为美洛培南+红霉素+克林霉素+复方新诺明+四环素(21.16%);青霉素敏感菌株和青霉素不敏感菌株相比,红霉素、阿莫西林、头孢吡肟、头孢噻肟、美洛培南、复方新诺明、克林霉素不敏感率差异有统计学意义(P<0.05)。结论:重庆地区红霉素、四环素、克林霉素耐药率较高,不宜用于儿童SP感染的治疗;青霉素和头孢类抗生素不敏感率较高,应慎重用于经验治疗。临床应根据药敏试验结果合理用药,防止耐药菌株流行播散。