New design and optimization of an in-house quantitative TaqMan Real-Time PCR-based assay for the detection and monitoring of occult hepatitis B virus (genotype A-J) infection

PurposeHBV DNA quantification is used for individuals with uninterpretable serological tests, occult HBV infections, decreasing the window period of the disease, and treatment follow-up. Although there are commercial qPCR assays, they are expensive. In this study, we developed a highly sensitive quantitative TaqMan Real-Time PCR with an exogenous internal control to quantify HBV DNA in serum/plasma.MethodsA specific primer/probe set was designed for the S conserved region of various HBV genotypes. The primer/probe set was evaluated experimentally and in-silico. An exogenous internal control was included to monitor the effects of inhibitors. The standard plasmid was titrated using three different methods to prepare the seven standards for the assay. The functional characteristics of the in-house assay were evaluated using the standards. Two hundred clinical specimens were also tested.ResultsThe LOD of the in-house assay was 40 IU/mL, and the assay was linear from 3.26Log10 to 9.26Log10 IU/mL. The analytical and clinical sensitivity of the assay was 100% and 92.15%, respectively. The analytical and clinical specificity of the assay was 100% and 98.97%, respectively. The positive and negative predictive values of the assay were determined to be 98.94% and 92.38%, respectively. The highest coefficient of variation of the inter/intra-assay was 5.1%. The accuracy was close to 100% for all standards, and the correlation between the in-house assay and commercial kit AltoStar® PCR Kits 1.5 was remarkable. The results of the clinical samples using the standards titrated using AcroMetrix? HBV Panel, Artus® HBV RG PCR Kit, and AltoStar® PCR Kits 1.5 were comparable (r ?= ?0.942, 0.951, 0.951).ConclusionsThe results indicate that the in-house assay is highly sensitive and specific, reproducible, and cost-benefit. Thus, it can be used to detect and quantify HBV DNA in research and clinical settings.     

Lower mean platelet volume is a risk indicator of hepatocellular carcinoma recurrence following liver transplantation

BackgroundLower mean platelet volume (MPV) is an indicator of platelet activity in the setting of tumor development. This study was to assess the relationship between preoperative MPV and survival outcomes of patients with hepatocellular carcinoma (HCC) following liver transplantation (LT).MethodsThe demographic and clinical characteristics of 304 HCC patients following LT were retrieved from an LT database. All the patients were divided into the normal and lower MPV groups according to the median MPV. The factors were first analyzed using a Kaplan–Meier survival analysis, then the factors with P < 0.10 were selected for multivariate Cox regression analysis and were used to define the independent risk factors for poor prognosis.ResultsThe 1-, 3-, and 5-year tumor free survival was 95.34%, 74.67% and 69.29% in the normal MPV group, respectively, and 95.40%, 59.97% and 42.94% in the lower MPV group, respectively (P < 0.01). No significant difference was observed in post-LT complications between the normal and lower MPV groups. Portal vein tumor thrombosis (PVTT) [hazard ratio (HR = 2.24; 95% confidence interval: 1.46–3.43; P < 0.01) and lower MPV (HR = 1.58; 95% confidence interval: 1.05–2.36; P = 0.03) were identified as independent prognostic risk factors for recipient survival.ConclusionPreoperative lower MPV is a risk indicator of HCC patients survival outcomes after LT.     

两种矫形方法治疗成人特发性脊柱侧凸的疗效比较

目的:比较单纯后路矫形术和一期前路松解、Halo-股骨髁上牵引加二期后路矫形术治疗成人特发性脊柱侧凸的疗效。方法:选取我院脊柱外科2003年1月～2007年12月收治的有完整影像学资料、Cobb角65°～90°的成人特发性脊柱侧凸患者30例,年龄20～30岁,平均23.4岁。均为初次手术,术前无神经损害。根据不同手术方法分为两组,行单纯后路矫形术的14例患者为A组,行一期前路松解、Halo-股骨髁上牵引及二期后路矫形术的16例患者为B组。两组患者术前侧凸Cobb角、胸椎后凸角、年龄、性别比、侧凸类型相匹配。随访时间为12～72个月,平均40个月。比较两组患者手术时间、出血量、住院时间、并发症情况、侧凸矫正率和冠状面平衡情况。结果:平均手术时间和平均住院时间A组分别为6.7±1.2h和24±18d,B组分别为9.9±1.4h和41±10d,B组均显著长于A组(P<0.05)。所有病例术后均无瘫痪、呼吸衰竭、死亡等并发症发生。术后侧凸矫正率A组为(51.3±11.8)%,B组为(64.5±11.6)%,B组显著大于A组(P<0.05);胸椎后凸角、C7中垂线与骶骨中线的距离A组为20.6°±8.4°、1.32±0.65cm,B组为20.4°±6.7°、1.30±0.70cm,两组比较均无显著性差异(P>0.05)。末次随访时A组侧凸矫正丢失率为(3.5±2.4)%,B组为(2.8±1.5)%,两组无显著性差异(P>0.05)。结论:两种治疗方案治疗中度成人特发性脊柱侧凸均可获得较好的畸形矫正,一期前路松解、Halo-双侧股骨髁上牵引可以增加侧凸Cobb角矫正率,但是存在显著增加手术时间和住院时间等不足。     

全椎弓根螺钉与钉钩联合系统治疗青少年Chiari畸形伴胸椎侧凸的疗效比较

 目的 评估青少年Chiari畸形伴胸椎侧凸不同内固定模式的选择对手术疗效的影响。 方法 回顾性分析2001年3月至2011年3月期间,接受后路胸椎融合术的75例Chiari畸形伴脊柱侧凸患者的病历资料,根据内固定方式分为全椎弓根螺钉组(44例)和钉钩联合组(31例)。分别测量术前、术后以及末次随访时的影像学指标：冠状面侧凸Cobb角、侧凸柔韧性、顶椎偏移、顶椎旋转及躯干偏移;矢状面胸椎后凸角、腰椎前凸角、躯干偏移及近端与远端交界区Cobb角。比较两组术前、术后及末次随访时上述影像学指标的改变。根据术前不同程度胸椎后凸进一步分组,比较两种内固定模式的手术疗效。结果 全椎弓根螺钉组术后胸弯平均矫正率为60.2％,明显著高于钉钩联合组(51.3％,t=2.372,P=0.023)。末次随访时全椎弓根螺钉组及钉钩联合组胸弯矫正丢失率分别为0.3％及1.7％(t=-0.468,P >0.05)。术后腰弯平均矫正率在全椎弓根螺钉组为61.7％,明显优于钉钩联合组51.1％(t=2.431,P=0.020)。术前全椎弓根螺钉组与钉钩联合组的胸弯顶椎偏移分别平均为25.0 mm和24.1 mm,术后减小至6.9 mm和7.4 mm,两组术后的胸弯顶椎偏移均获得明显改善。术前胸椎后凸>40°的病例中,全椎弓根螺钉组末次随访时矢状面近端交界区Cobb角为10.0°,高于钉钩联合组(4.5°,t=-2.031,P=0.052)。而且全椎弓根螺钉组近端交界性后凸发生率(20% )高于钉钩联合组(9%)。结论 对继发于青少年Chiari畸形的胸椎侧凸行后路内固定矫形,全椎弓根螺钉具有更好的畸形矫正率,但是,与钉钩联合固定相比,术前胸椎过度后凸的患者在胸椎全椎弓根螺钉固定术后远期发生近端交界性后凸的风险增高。     

益生菌Faecalibacterium prausnitzii联合溃克灵对大鼠结肠炎的预防作用

目的:探讨益生菌 Faecalibacterium prausnitzii(F. prausnitzii)联合溃克灵防治炎症性肠病的疗效及作用机制?方法:将50只Sprague-Dawley大鼠随机分为5组:正常组?模型组?F.prausnitzii组?溃克灵组?F.prausnitzii联合溃克灵组?造模前各组灌胃1周(正常组和模型组予生理盐水,其余3组给予相应的F.prausnitzii和溃克灵处理)?采用2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)法(80 mg/kg TNBS联合等体积无水乙醇)灌肠复制出结肠炎大鼠模型,3 d后全部处死,观察结肠组织损伤情况,行结肠组织病理学评分;ELISA 法测定各组大鼠血浆白介素(interleukin,IL)-10?IL-12?肿瘤坏死因子(tumor necrosis factor,TNF)-α?干扰素(interferon,IFN)-γ水平,计算IL-10/IL-12比值;免疫组化检测肠黏膜IL-10及TNF-α的表达?结果:F.prausnitzii联合溃克灵组与模型组相比,结肠组织损伤明显改善,组织病理学评分显著降低(P < 0.05),血浆IL-10?IL-10/IL-12比值显著升高(P均 < 0.05),血浆IL-12?IFN-γ显著下降(P < 0.05,P < 0.001),血浆TNF-α下降但差异无统计学意义,肠黏膜IL-10表达明显升高(P < 0.01),肠黏膜TNF-α表达明显降低(P < 0.05)?与F.prausnitzii组?溃克灵组相比,F.prausnitzii联合溃克灵组大鼠结肠组织病理学评分降低,IL-10/IL-12比值及血浆IL-10?肠黏膜IL-10表达升高,血浆IL-12?IFN-γ?TNF-α及肠黏膜TNF-α表达下降,但差异无统计学意义?结论:益生菌F.prausnitzii联合中药溃克灵对结肠炎大鼠肠黏膜的预防保护作用,可能与促进外周血及肠黏膜IL-10分泌从而抑制外周血IL-12?TNF-α?IFN-γ产生?降低肠黏膜TNF-α表达等免疫调节机制有关?     

尿毒清颗粒调控TGF-β1/SnoN/Smads信号通路改善肾衰竭模型鼠肾间质纤维化的作用和机制

为了初步阐明尿毒清颗粒(uremic clearance granule,UCG)在体内调控转化生长因子(transforming growth factor,TGF)-β1/SnoN/Smads信号通路而改善肾间质纤维化(renal interstitial fibrosis,RIF)的作用和机制,将15只大鼠随机分为正常组、模型组、尿毒清组。采用腺嘌呤灌胃联合单侧输尿管结扎术(unilateral ureteral obstruction,UUO)建立肾衰竭模型。造模后,3组大鼠分别给予UCG悬浊液或蒸馏水,共3周,其间,检测大鼠体重、24 h尿蛋白排泄量(urinary protein excretion,Upro);给药3周后,处死全部大鼠,抽取血液,摘除双肾,称重,观察肾脏外观和肾组织形态特征,检测血清生化指标和肾组织TGF-β1,SnoN,磷酸化Smad2/3(phosphorylated Smad2/3,p-Smad2/3)以及Smad7蛋白表达量。结果表明,经UCG干预后,模型鼠一般情况,肾脏外观、血清肌酐(serum creatinine,Scr)、血清尿素氮(blood urea nitrogen,BUN)、尿酸(uric acid,UA)、白蛋白(albumin,Alb)、Upro以及肾组织形态均得到不同程度的改善;UCG还可以下调模型鼠肾组织TGF-β1,p-Smad2/3蛋白表达水平,上调SnoN,Smad7蛋白表达水平。总之,UCG可能在体内多靶点地调控TGF-β1/SnoN/Smad信号通路,从而,减少细胞外基质(extracellular matrix,ECM)合成,延缓肾衰竭进展。     

Nutritional assessment and risk factors associated to malnutrition in patients with esophageal cancer

Introduction: Esophageal cancer is the fourth most common cause of cancer death in China. Patients with esophageal cancer are more likely to suffer from malnutrition. The purpose of this study is to assess nutritional status of patients with esophageal cancer from multiple perspectives and analyze the risk factors. Methods: A total of 1482 esophageal cancer patients were enrolled in the study. We investigated the Scored Patient Generated Subjective Global Assessment (PG-SGA) scores, NRS-2002 scores, Karnofsky performance status scores, anthropometric, and laboratory indicators of patients. Unconditional logistic regression analysis was applied to identify the risk factors of nutritional status. Results: PG-SGA (≥4) and NRS-2002 (≥3) showed the incidence of malnutrition were 76% and 50%, respectively. In the patients with PG-SGA score ≥4, the proportion of patients who did not receive any nutritional support was 60%. The incidence of malnutrition in females was significantly higher than that in males. Besides, abnormality rates of Red blood cell (P < 0.001), MAC (P = 0.037), and MAMC (P < 0.001) in males was significantly higher than that in females, while abnormality rates of TSF (P < 0.001) was lower than that in females. After adjusted with the other potential risk factors listed, unconditional logistic regression analysis indicated smoking (odds ratio: 2.868, 95% confidence interval: 1.660-4.954), drinking (OR: 1.726, 95% CI: 1.099-2.712), family history (OR: 1.840, 95% CI: 1.132-2.992), radiotherapy or chemotherapy (OR: 1.594, 95% CI: 1.065-2.387), and pathological stage (OR: 2.263, 95% CI: 1.084-4.726) might be the risk factors of nutritional status, while nutritional support can reduce the risk of malnutrition. Conclusion: Effective nutritional risk assessment methods and nutritional intervention measures can be adopted according to the research data to improve quality of life of esophageal cancer patients.