The downregulation of ANGPTL4 inhibits the migration and proliferation of tongue squamous cell carcinoma

ObjectiveTongue squamous cell carcinoma (TSCC) is the most common malignant cancer in the oral cavity, with a high rate of metastasis to the neck lymphoid node. Angiopoietin-like protein 4 (ANGPTL4) and microvessel density (MVD) may be novel indicators for tumor metastasis. The aim of the present study was to investigate the expression and function of ANGPTL4 in TSCC and the relationship between ANGPTL4 and MVD.MethodsThe expression levels of ANGPTL4 and MVD (CD34) were analyzed in 65 TSCC specimens and the adjacent non-cancerous tissues using immunohistochemistry (IHC). siRNA was delivered into TSCCA cells to downregulate ANGPTL4 expression. Subsequently, validation with real-time RT-PCR and western blot analyses was performed to analyze ANGPTL4 expression levels. In addition, a proliferation assay, migration and invasion assays were carried out.ResultsANGPTL4 expression was associated with tumor stage, lymph node metastasis and MVD expression. Cox regression analysis showed that high levels of ANGPTL4 expression were closely associated with poor survival time. In vitro analyses using qRT-PCR and western blot confirmed that ANGPTL4 was successfully inhibited in TSCCA cells. Suppressing ANGPTL4 resulted in the inhibition of cell proliferation and migration, but neither invasion nor cisplatin resistance was significantly affected.ConclusionHigh expression levels of ANGPTL4 are associated with the T stage, lymphatic metastasis, angiogenesis and poor overall survival in TSCC patients. The downregulation of ANGPTL4 inhibits the migration and proliferation of cells in TSCC. Taken together, ANGPTL4 may serve as a novel biomarker and therapeutic target for TSCC.     

鱼龙草颗粒止咳祛痰抗炎作用实验研究

目的 探讨鱼龙草颗粒对小鼠的止咳、祛痰、抗炎作用,阐明鱼龙草颗粒药效学作用机制,为临床使用提供理论依据.方法 将150只小鼠（做3个试验,每个试验50只,每个试验分5组,每组10只）随机分为空白对照组（去离子水,0.2 ml/只）、阳性药物组（磷酸可待因30 mg/kg或盐酸氨溴索0.05 g/kg或阿司匹林0.2 g/kg）和观察组（鱼龙草颗粒）中高剂量组（12.0 g/kg）、中剂量组（6.0 g/kg）、低剂量组（3.0 g/kg）三剂量组,按分组剂量每天上午9：00左右,每只小鼠灌胃给药1次,连续给药5d后进行实验：①小鼠SO2引咳试验：用SO2刺激小鼠呼吸道15 s引起小鼠咳嗽反应,观察3种不同剂量组及磷酸可待因组小鼠咳嗽潜伏期及咳嗽次数;②小鼠气管酚红分泌排泄试验：利用酚红注射后可在小鼠气管分泌排泄机制,腹腔注射2.5％酚红氯化钠注射液30 min,空气栓塞处死小鼠,取出气管,测量3种不同剂量组及盐酸氨溴索组小鼠气管对酚红分泌排泄量;③小鼠二甲苯所致耳廓肿胀试验：利用二甲苯外涂可引起小鼠耳廓炎症反应方法,给予小鼠右耳涂抹二甲苯（100％浓度）,4h后处死小鼠,取下左右耳,对3种不同剂量组及阿司匹林组称重.以上3种试验均设空白对照组并与3种不同剂量组及阳性药物组进行对比观察.结果 ①小鼠SO2引咳试验结果显示：在延长小鼠咳嗽潜伏期方面,高、中剂量组与空白组比较,差异有统计学意义[（60±17）、（58±15）s比（50±18）s] （P ＜0.05）;在减少小鼠咳嗽次数方面,高、中剂量组与空白组比较,差异有统计学意义[（15±5）、（16±9）次/3 min比（35±10）次/3 min] （P ＜0.01）,与低剂量组[（20±7）次/3 min]比较差异有统计学意义（P＜0.05）;磷酸可待因组[咳嗽潜伏期（68±15）s、咳嗽次数（12±6）次/3 min]与空白组比较,差异有统计学意义（P＜0.01）.证明鱼龙草颗粒可延长小鼠咳嗽潜伏期、明显减少咳嗽次数.②空白对照组、鱼龙草颗粒高中低剂量组、盐酸氨溴索组酚红量分别为（1.03±0.37）、（1.59±0.23）、（1.56±0.21）、（1.43±0.26）、（1.55±0.22） mg/L.小鼠气管酚红分泌排泄试验结果显示：高、中剂量组与空白组比较,差异有统计学意义（P＜0.01）;低剂量组与空白组比较,差异有统计学意义（P＜0.05）;盐酸氨溴索组与空白组比较,差异有统计学意义（P＜0.01）.③小鼠结果显示：高、中、低3种剂量组及阿司匹林组4组与空白组比较,差异均有统计学意义[（8.2±2.0）、（10.0±1.1）、（10.9±1.6）、（8.0±1.1）mg比（14.2±1.7）mg]（均P＜0.01）.结论 小鼠的SO2引咳试验、气管酚红分泌排泄试验、二甲苯所致耳廓肿胀试验显示：鱼龙草颗粒对小鼠具有明显的止咳、祛痰、抗炎作用,其临床疗效有待进一步观察.     

高效毛细管电泳法测定黄连中盐酸小檗碱的含量

目的建立高效毛细管电泳法(HPCE)测定黄连中盐酸小檗碱的含量。方法熔融石英毛细管柱50 cm×75μm,缓冲液为0.05 mol.L-1四硼酸钠溶液(pH=7.0)-甲醇(85∶15),分离电压14 kV,毛细管柱温20℃,UV检测波长254 nm。结果盐酸小檗碱进样浓度在0.026~0.414 mg.mL-1内线性关系良好(r=0.999 4),平均加样回收率为98.90%(RSD=2.64%,n=5)。结论本法灵敏、快速、准确,可用于黄连的质量控制。     

Development of a machine learning algorithm to predict complications of total laparoscopic anterior resection and natural orifice specimen extraction surgery in rectal cancer

BackgroundTotal laparoscopic anterior resection (tLAR) and natural orifice specimen extraction surgery (NOSES) has been widely adopted in the treatment of rectal cancer (RC). However, no study has been performed to predict the short-term outcomes of tLAR using machine learning algorithms to analyze a national cohort.MethodsData from consecutive RC patients who underwent tLAR were collected from the China NOSES Database (CNDB). The random forest (RF), extreme gradient boosting (XGBoost), support vector machine (SVM), deep neural network (DNN), logistic regression (LR) and K-nearest neighbor (KNN) algorithms were used to develop risk models to predict short-term complications of tLAR. The area under the receiver operating characteristic curve (AUROC), Gini coefficient, specificity and sensitivity were calculated to assess the performance of each risk model. The selected factors from the models were evaluated by relative importance.ResultsA total of 4313 RC patients were identified, and 667 patients (15.5%) developed postoperative complications. The machine learning model of XGBoost showed more promising results in the prediction of complication than other models (AUROC 0.90, P < 0.001). The performance was similar when internal and external validation was used. In the XGBoost model, the top four influential factors were the distance from the lower edge of the tumor to the anus, age at diagnosis, surgical time and comorbidities. In risk stratification analysis, the rate of postoperative complications in the high-risk group was significantly higher than in the medium- and low-risk groups (P < 0.001).ConclusionThe machine learning model shows potential benefits in predicting the risk of complications in RC patients after tLAR. This novel approach can provide reliable individual information for surgical treatment recommendations.     

5μg和10μg重组乙型肝炎疫苗初次免疫正常应答和高应答新生儿5年抗体持久性比较

目的 比较5 μg和10 μg乙型肝炎疫苗（HepB）初次免疫（初免）正常应答和高应答新生儿初免后5年的抗体持久性。方法 选用5 μg（重组啤酒酵母）和10 μg（重组汉逊酵母）HepB,按照"0-1-6"程序完成3剂次初免的新生儿,并在接种第3剂次后1~6个月（T₀）和5年（T₁）后分别采集静脉血,采用化学发光微粒子免疫分析法（CMIA）定量检测抗-HBs,比较两剂量接种后正常应答和高应答（T₀时抗-HBs ≥ 100 mIU/ml）者T₁时的抗体阳性率（抗-HBs ≥ 10 mIU/ml）和平均抗体浓度（GMC）;通过多因素分析探讨接种剂量与抗体持久性的关系。结果 HepB 5 μg组和10 μg组分别共有1 883名和1 495名观察对象纳入分析,T₁时抗-HBs阳性率分别为49.92%（943/1 883）和75.92%（1 135/1 495）,差异有统计学意义（χ²=237.75,P<0.001）;GMC分别为10.23（95%CI：9.38~11.16）mIU/ml和28.91（95%CI：26.65~31.35）mIU/ml,差异有统计学意义（F=280.36,P<0.001）。10 μg组T₁时抗-HBs阴转者抗-HBs滴度分布与5 μg组的差异有统计学意义（χ²=39.75,P<0.001）。多因素分析显示,排除其他因素影响后,HepB初免剂量与T₁时抗-HBs阳性率和抗-HBs滴度均独立相关[P<0.001,OR=1.44（95%CI：1.20~1.73）;P<0.001,β=0.27（95%CI：0.14~0.40）]。结论 新生儿使用10 μg重组HepB初免后5年抗-HBs持久性优于5 μg重组HepB。     

Vacuolating cytotoxin purified fromHelicobacter pyloricauses mitochondrial damage in human gastric cells

We investigated the effects of vacuolating cytotoxin (VacA) prepared fromHelicobacter pylorion the metabolism of gastric epithelial cells, AZ-521. VacA caused the ATP levels to decrease in a time-dependent manner; by approximately 20% in 6 h, 35% in 12 h and 50% in 24 h, at a concentration of 120 nM. This decrease was also dependent on the concentration of VacA. To evaluate the impairment of mitochondria by VacA, mitochondrial membrane potential was estimated by flow cytometric analysis using 3, 3′-dihexyloxacarbocyanine iodide as a substrate. VacA decreased membrane potential with the relative fluorescence intensity of AZ-521 cells in 6 h from 52±3 to 24±1. Treatment of the cells with bafilomycin A1, a specific inhibitor of vacuolar ATPase proton pump, showed no apparent effect on these changes in the levels of ATP and the mitochondrial membrane potential. Secondly, we estimated the effect of VacA on oxygen consumption. VacA inhibited oxygen consumption in AZ-521 cells: the levels of PO₂in the medium of control cells decreased by 73% in 3 h and 37% in 6 h, whereas those in VacA-treated cells were 84% in 3 h and 59% in 6 h. Flow cytometric analysis showed the number of cells in the G₀/G₁phase was increased by VacA. Taken together, VacA induced an inactivation of energy metabolism followed by mitochondrial damage, leading to impairment of the cell cycle in gastric epithelial cells.     

Validation of Two Clinical Prognostic Models in Patients With Acute Symptomatic Pulmonary Embolism

ObjectiveThe aim of this study was to compare the predictive value of two clinical prognostic models, the Spanish score and the simplified Pulmonary Embolism Severity Index (sPESI), in an independent cohort of patients diagnosed of acute symptomatic pulmonary embolism (PE).MethodsWe performed a retrospective analysis of a cohort composed of 1447 patients with acute symptomatic PE. The Spanish score and the sPESI were calculated for each patient according to different clinical variables. We assessed the predictive accuracy of these scores for 30-day mortality, and a composite of non fatal recurrent venous thromboembolism and non fatal major bleeding, using C statistic, which was obtained by means of logistic regression and ROC curves.ResultsOverall, 138 patients died (9.5%) during the first month of follow-up. Both scores showed an excellent predictive value for 30-day all-cause mortality (C statistic, 0.72 and 0.74), but the performance was poor for the secondary endpoint (C statistic, 0.60 and 0.59). The sPESI classified fewer patients as low risk (32% versus 62%; P<.001). Low-risk patients based on the sPESI had a lower 30-day mortality than those based on the Spanish score (1.1% versus 4.2%), while the 30-day rate of non fatal recurrent VTE or major bleeding was similar (2.2% versus 2.3%).ConclusionsBoth scores provide excellent information to stratify the risk of mortality in patients treated of PE. The usefulness of these models for nonfatal adverse events is questionable. The sPESI identified low-risk patients with PE better than the Spanish score.