α-细辛醚、β-细辛醚改善Aβ 25-35 诱导的 PC12细胞损伤及机制

目的：探究α-细辛醚、β-细辛醚对β淀粉样蛋白活性片段Aβ _25-35诱导的PC12细胞损伤模型的保护作用及相关作用机制。 方法：采用Aβ _25-35诱导PC12细胞建立Aβ毒性损伤细胞模型。将PC12细胞分为空白对照组、模型对照组、α-细辛醚组（0.5、1.0、1.5 μg/mL）、β-细辛醚组（6.3、12.5、25.0 μg/mL）、血管活性肠肽（VIP）组，并设VIP拮抗剂对照。采用细胞计数试剂盒（CCK-8）法检测细胞存活率；流式细胞术检测细胞凋亡率；酶联免疫吸附试验检测炎症因子白介素（IL）-1、IL-10、肿瘤坏死因子（TNF）-α，氧化因子诱生型一氧化氮合酶（iNOS）、一氧化氮（NO）和凋亡因子caspase-3、p53水平；蛋白质印迹法检测细胞c-Jun氨基端激酶（JNK）、p38丝裂原活化蛋白激酶（p38MAPK）蛋白表达。 结果：与模型对照组比较，α-细辛醚组、β-细辛醚组和VIP组细胞存活率增加，细胞凋亡率下降，凋亡因子caspase-3、p53和炎症因子IL-1、TNF-α水平下降，IL-10水平升高，氧化因子iNOS和NO水平下降，c-Jun氨基端激酶（JNK）、p38MAPK蛋白表达减少（均 P<0.05）。VIP拮抗剂干预后，β-细辛醚组细胞存活率下降，细胞凋亡率增加，凋亡因子caspase-3、p53和炎症因子IL-1、TNF-α水平升高，IL-10水平下降，氧化因子iNOS和NO水平升高，JNK、p38MAPK蛋白表达增加（均 P<0.05）；α-细辛醚组无显著变化（均 P>0.05）。 结论：α-细辛醚、β-细辛醚对Aβ _25-35诱导的PC12细胞损伤模型具有保护作用，β-细辛醚可通过促进VIP的分泌，调控JNK/MAPK通路从而抑制炎症因子、氧化因子水平，改善PC12细胞凋亡；α-细辛醚作用机制与VIP分泌水平无明显联系。     

Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon α2b

Recombinant human interferon α2b (rhIFNα2b) is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C. The current identification test for rhIFNα2b is complex. In this study, an anti-rhIFNα2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNα2b. RhIFNα2b was used to immunize an alpaca, which established a phage nanobody library. After five steps of enrichment, the nanobody I22, which specifically bound rhIFNα2b, was isolated and inserted into the prokaryotic expression vector pET28a. After subsequent purification, the physicochemical properties of the nanobody were determined. A semiquantitative detection and rapid identification assay of rhIFNα2b was developed using this novel nanobody. To develop a rapid test, the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips. The developed rhIFNα2b detection assay had a limit of detection of 1 μg/mL. The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products. The principle of this novel assay is generally applicable for the rapid testing of other commercial products, with a great potential for routine use in detecting counterfeit recombinant protein products.     

Impact of thymosin α1 as an immunomodulatory therapy on long-term survival of non-small cell lung cancer patients after R0 resection: a propensity score-matched analysis

Background:There is limited information about thymosin α1 (Tα1) as adjuvant immunomodulatory therapy, either used alone or combined with other treatments, in patients with non-small cell lung cancer (NSCLC). This study aimed to evaluate the effect of adjuvant Tα1 treatment on long-term survival in margin-free (R0)-resected stage IA–IIIA NSCLC patients.Methods:A total of 5746 patients with pathologic stage IA-IIIA NSCLC who underwent R0 resection were included. The patients were divided into the Tα1 group and the control group according to whether they received Tα1 or not. A propensity score matching (PSM) analysis was performed to reduce bias, resulting in 1027 pairs of patients.Results:After PSM, the baseline clinicopathological characteristics were similar between the two groups. The 5-year disease-free survival (DFS) and overall survival (OS) rates were significantly higher in the Tα1 group compared with the control group. The multivariable analysis showed that Tα1 treatment was independently associated with an improved prognosis. A longer duration of Tα1 treatment was associated with improved OS and DFS. The subgroup analyses showed that Tα1 therapy could improve the DFS and/or OS in all subgroups of age, sex, Charlson Comorbidity Index (CCI), smoking status, and pathological tumor-node-metastasis (TNM) stage, especially for patients with non-squamous cell NSCLC and without targeted therapy.Conclusion:Tα1 as adjuvant immunomodulatory therapy can significantly improve DFS and OS in patients with NSCLC after R0 resection, except for patients with squamous cell carcinoma and those receiving targeted therapy. The duration of Tα1 treatment is recommended to be >24 months.     

Hepatocyte nuclear factor 4α in the pathogenesis of non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is emerging as the most common chronic liver disease worldwide. It refers to a range of liver conditions affecting people who drink little or no alcohol. NAFLD comprises non-alcoholic fatty liver and non-alcoholic steatohepatitis (NASH), the more aggressive form of NAFLD. NASH is featured by steatosis, lobular inflammation, hepatocyte injury, and various degrees of fibrosis. Although much progress has been made over the past decades, the pathogenic mechanism of NAFLD remains to be fully elucidated. Hepatocyte nuclear factor 4α (HNF4α) is a nuclear hormone receptor that is highly expressed in hepatocytes. Hepatic HNF4α expression is markedly reduced in NAFLD patients and mouse models of NASH. HNF4α has been shown to regulate bile acid, lipid, glucose, and drug metabolism. In this review, we summarize the recent advances in the understanding of the pathogenesis of NAFLD with a focus on the regulation of HNF4α and the role of hepatic HNF4α in NAFLD. Several lines of evidence have shown that hepatic HNF4α plays a key role in the initiation and progression of NAFLD. Recent data suggest that hepatic HNF4α may be a promising target for treatment of NAFLD.     

Use of unidirectional and spherical porous β-tricalcium phosphate in opening-wedge high tibial osteotomy: a case series

Introduction: Unidirectional porous β-tricalcium phosphate (UDPTCP) consists of a novel porous artificial bone that is structurally different from conventional artificial bone comprised of spherical porous β-tricalcium phosphate (SPTCP).Case presentation: We present our first four clinical cases of opening-wedge high tibial osteotomy (OWHTO) using UDPTCP and SPTCP together. The patients’ mean age was 54.5 ± 5.9 years, and the mean observation period was 20.8 ± 2.8 months. In OWHTO, two wedge shaped pieces of UDPTCP and SPTCP were cut to fit the gap and implanted parallel to each other in the anterior and posterior parts, respectively. We evaluated the correction loss and bone remodeling for UDPTCP and SPTCP over time using radiography and computed tomography, and evaluated the clinical outcomes.Conclusion: There was no correction loss reported in any case, and early bone remodeling was observed with UDPTCP. All patients achieved satisfactory clinical results with no adverse events.     

皮肤鳞状细胞癌和基底细胞癌组织中VEGF、COX-2及ID-1的表达及临床意义

目的 研究皮肤鳞状细胞癌（SCC）和基底细胞癌（BCC）组织中血管内皮生长因子（VEGF）、环氧化酶-2（COX-2）及分化抑制因子-1（ID-1）的表达和临床意义。方法 选取2016年3月—2019年3月重庆市江津区中心医院收治的皮肤恶性肿瘤患者76例为研究对象,其中SCC组和BCC组均38例。配对选取30例无恶性肿瘤的同期皮肤移植患者为对照组。采用免疫组织化学法检测VEGF、COX-2及ID-1的阳性表达率,RT-PCR检测mRNA的表达,Western blotting法检测蛋白相对表达量,并分析其相关性。结果 3组VEGF的阳性表达率、mRNA及蛋白表达的比较,差异有统计学意义（P?<0.05）;SCC组与BCC组高于对照组（P?<0.05）;SCC组高于BCC组（P?<0.05）。VEGF、COX-2及ID-1的表达均呈正相关（P?<0.05）。结论 皮肤SCC与BCC的产生和进展与VEGF、COX-2及ID-1的高表达有关,且VEGF、COX-2及ID-1的表达均呈正相关。联合检测VEGF、COX-2及ID-1的表达对皮肤SCC与BCC的早期诊治具有重要意义。     

吡非尼酮通过下调TGF-β/Smad通路中TGF-β3的表达抑制兔Tenons囊成纤维细胞增殖

目的探讨吡非尼酮在抗青光眼滤过术后抑制瘢痕形成的可能机制。方法体外培养兔眼Tenons囊成纤维细胞(RYTF)；设计6组不同浓度的吡非尼酮作为实验组、1组未加入吡非尼酮的作为对照组及1组调零组，采用CCK-8检测法初步确定吡非尼酮的起始作用浓度及最适作用浓度；采用免疫荧光法检测经起始、最适作用浓度吡非尼酮作用后RYTF中TGF-β3，Ⅰ、Ⅲ型胶原蛋白与对照组相比的荧光染色表达情况；通过Western blot法检测经起始、最适作用浓度吡非尼酮作用后TGF-β3，CollagenⅠ、CollagenⅢ因子与对照组相比的蛋白表达情况；通过RT-PCR法检测经起始、最适作用浓度吡非尼酮作用后TGF-β3、CollagenⅠ、CollagenⅢ与对照组相比的mRNA表达变化。结果吡非尼酮抑制RYTF增殖的起始作用浓度和最适作用浓度分别为0.1 mg和0.27 mg；与对照组相比，经起始、最适作用浓度吡非尼酮作用24 h后，实验组RYTF中TGF-β3、CollagenⅠ、Collagen Ⅲ的荧光表达均少于对照组（P < 0.05）；与对照组相比，经起始、最适作用浓度吡非尼酮作用24 h后，实验组RYTF中TGF-β3、CollagenⅠ、CollagenⅢ的蛋白表达量均少于对照组（P < 0.05）；与对照组相比，经起始、最适作用浓度吡非尼酮作用24h后，实验组RYTF中TGF-β3、CollagenⅠ、Collagen Ⅲ的基因相对表达量均少于对照组（P < 0.05）。结论吡非尼酮对体外培养RYTF增殖有明显抑制作用，且呈浓度依赖关系；吡非尼酮抑制RYTF增殖的机制可能是通过影响RYTF中TGF-β/Smad途径上TGF-β3效应因子的基因和蛋白表达。     

Serum S100β as a predictor of severity and outcomes for mixed subtype acute ischaemic stroke

INTRODUCTIONSerum S100β levels are mostly used for predicting outcomes of large-vessel stroke. Its application to mixed subtypes of acute ischaemic stroke (AIS) has been limited.METHODSPatients with mixed subtypes of AIS who were aged over 18 years and presented within 24 hours of stroke onset were consecutively enrolled. Serum S100β levels at presentation (S100β_b) and 72 hours (S100β_72hrs), and corresponding National Institutes of Health Stroke Scale (NIHSS_b and NIHSS_72hrs, respectively) scores were assessed. Stroke outcomes were evaluated using the modified Rankin Scale (mRs) at 30 days (mRs₃₀) and 90 days (mRs₉₀). Correlations between S100β_b and S100β_72hrs, as well as differences between the two (∆S100β) and the corresponding NIHSS, mRs₃₀ and mRs₉₀ scores, were evaluated (p < 0.05).RESULTS35 patients were eligible for analysis. On univariate analysis, stroke outcomes had a significant association with S100β_b, S100β_72hrs, NIHSS_b, NIHSS_72hrs and ∆S100β. Both S100β_b and S100β_72hrs correlated with corresponding NIHSS values (ρ_b = 0.51, p < 0.001; ρ_72hrs = 0.74, p < 0.001), mRs₃₀ (ρ_b = 0.58, p < 0.001; ρ_72hrs = 0.72, p < 0.001) and mRs₉₀ (ρ_b = 0.51, p = 0.002; ρ_72hrs = 0.68, p < 0.001). Correlations existed between ∆S100β and mRs₃₀ (ρ = 0.74, p < 0.001) and mRs₉₀ (ρ = 0.71, p < 0.001). Practical cut-off points for unfavourable outcomes (mRs 3–6) were S100β_72hrs > 0.288 µg/L (sensitivity 92.3%, specificity 86.4%) and ∆S100β > 0.125 µg/L (sensitivity 100%, specificity 81.8%).CONCLUSIONHigh serum S100β is associated with unfavourable outcomes for mixed subtype AIS. Cut-off values of S100β_72hrs and ∆S100β were optimal for predicting unfavourable stroke outcomes.     

Mir-106b表达与食管鳞状细胞癌的关系

目的研究微小RNA106b（mir-106b）在食管鳞状细胞癌组织中的表达情况,并进一步分析其与肿瘤的临床病理学特征及 预后的相关性。方法收集200例2001~2007年中国医学科学院接收的食管鳞状细胞癌患者手术切除获得的肿瘤组织和相对应 的癌旁组织,采用qRT-PCR方法检测mir-106b在组织中的相对表达水平,并通过northern blot进一步验证其表达情况。应用统 计学方法分析mir-106b表达量与肿瘤临床病理学特征及预后的相关性。结果相对于癌旁组织,食管鳞状细胞癌肿瘤组织中 mir-106b的表达水平显著升高,统计学数据显示,mir-106b的表达量与淋巴结转移、肿瘤分期及吸烟有相关性（P<0.05）。并且, mir-106b表达水平较低的患者的生存率（60个月）显著高于mir-106b表达水平较高的患者（37个月,P=0.024）。另外,Cox回归 分析显示,mir-106b的表达量、区域淋巴结转移及吸烟均为是食管鳞状细胞癌患者独立预后因子（P<0.05）。结论mir-106b在 食管鳞状细胞癌患者肿瘤组织中高表达,且与淋巴结转移及预后不良密切相关,可用作ESCC的诊断及预后生物标志物。     

羟尼酮可阻止大鼠肝星状细胞活化：基于抑制TGF-β1通路蛋白的磷酸化

目的探讨羟尼酮对CCl4诱导的大鼠肝纤维化的作用及相关机制。方法66只雄性SD大鼠按随机数字法分为对照组10只、模型组20只、羟尼酮（100 mg/kg）组12只、羟尼酮（250 mg/kg）组12只、吡非尼酮（250 mg/kg）组12只，采用CCl₄皮下注射进行肝纤维化造模，羟尼酮、吡非尼酮予以灌胃给药，处死大鼠后，留取血清检测肝功能，利用肝脏组织检测羟脯氨酸的含量，肝组织进行HE染色和Sirius Red染色，对炎症和纤维化程度进行评分。将人肝星状细胞系LX-2分组：对照组、TGF-β1组、羟尼酮组、吡非尼酮组，经过TGF-β1刺激和羟尼酮、吡非尼酮处理后，Western blot分别检测α-SMA、I型胶原蛋白和磷酸化Smad3、磷酸化p38、磷酸化Erk1/2、磷酸化Akt等磷酸化蛋白。结果动物实验显示羟尼酮组大鼠肝功能值有改善，肝组织羟脯氨酸含量明显减少，肝组织纤维化程度显著降低，且均具有统计学意义（P < 0.05）。细胞实验发现随羟尼酮浓度的增高，ɑ-SMA和I型collagen蛋白表达水平降低；TGF-β1刺激细胞后，TGF-β信号转导通路中Smad3、P38、ERK、Akt等蛋白的磷酸化水平随着作用时间的延长而增加，但在相同作用时间下，羟尼酮组细胞TGF-β信号转导通路蛋白磷酸化的表达水平低于TGF-β1组细胞（P < 0.05）。结论羟尼酮可抑制TGF-β信号转导通路蛋白的磷酸化，从而阻止TGF-β1介导的肝星状细胞活化，可能是其缓解CCl₄诱导的大鼠肝纤维化机制之一。     

FUT8调控TGF-β1介导的肺成纤维细胞纤维化的机制

目的探讨α-1, 6岩藻糖基转移酶(FUT8)对人胚肺成纤维细胞(MRC-5)增殖、迁移和纤维化的作用及可能机制。方法将24只C57/BL6小鼠随机分为对照组、博莱霉素组、sh-NC组和sh-FUT8组, 应用Masson染色观察肺纤维化情况。细胞实验分别设置对照组: MRC-5正常培养; TGF-β1组: TGF-β1处理MRC-5;si-NC组: si-NC转染MRC-5后TGF-β1处理; si-FUT8组: si-FUT8转染MRC-5后TGF-β1处理; Gal-3组: si-FUT8和pcDNA3.1-Gal转染MRC-5后TGF-β1处理。CCK-8和BrdU方法检测细胞增殖; 划痕实验检测细胞迁移; RT-qPCR和Western blot检测α-平滑肌肌动蛋白(α-SM A)、I型胶原蛋白(COLIA1)和细胞外基质纤维连接蛋白(FN)的表达水平。同时, 免疫共沉淀检测了FUT8与半乳糖凝集素-3(Gal-3)的作用及沉默FUT8对FAK/Akt信号通路的调节。结果沉默FUT8显著降低博莱霉素诱导的小鼠肺组织细胞外胶原沉积。沉默FUT8抑制TGF-β1介导的细胞增殖(186.81±6.29 vs 118.09±9.48, P < 0.05)和迁移。FUT8缺失下调α-SMA、COLIA1和FN的mRNA和蛋白表达水平(P < 0.05)。此外, FUT8可直接与Gal-3作用。沉默FUT8下调Gal-3的表达并抑制FAK/Akt信号通路, 过表达Gal-3逆转FUT8对细胞增殖、迁移和纤维化的作用(P < 0.05)。结论FUT8通过调控Gal-3调控TGF-β1介导的MRC-5细胞增殖、迁移和纤维化, FAK/Akt信号通路可能参与了这个过程。     

Chronic hypoperfusion due to intracranial large artery stenosis is not associated with cerebral β-amyloid deposition and brain atrophy

Background:Insufficient cerebral perfusion is suggested to play a role in the development of Alzheimer disease (AD). However, there is a lack of direct evidence indicating whether hypoperfusion causes or aggravates AD pathology. We investigated the effect of chronic cerebral hypoperfusion on AD-related pathology in humans.Methods:We enrolled a group of cognitively normal patients (median age: 64 years) with unilateral chronic cerebral hypoperfusion. Regions of interest with the most pronounced hypoperfusion changes were chosen in the hypoperfused region and were then mirrored in the contralateral hemisphere to create a control region with normal perfusion. ¹¹C-Pittsburgh compound-positron emission tomography standard uptake ratios and brain atrophy indices were calculated from the computed tomography images of each patient.Results:The median age of the 10 participants, consisting of 4 males and 6 females, was 64 years (47–76 years). We found that there were no differences in standard uptake ratios of the cortex (volume of interest [VOI]: P = 0.721, region of interest [ROI]: P = 0.241) and grey/white ratio (VOI: P = 0.333, ROI: P = 0.445) and brain atrophy indices (Bicaudate, Bifrontal, Evans, Cella, Cella media, and Ventricular index, P > 0.05) between the hypoperfused regions and contralateral normally perfused regions in patients with unilateral chronic cerebral hypoperfusion.Conclusion:Our findings suggest that chronic hypoperfusion due to large vessel stenosis may not directly induce cerebral β-amyloid deposition and neurodegeneration in humans.     

宫内发育迟缓出生后追赶生长大鼠脂肪组织 LRP6/ β -catenin通路表达变化

目的：探讨宫内发育迟缓出生后追赶生长（CG-IUGR）大鼠脂肪组织低密度脂蛋白受体相关蛋白6（LRP6）/β-联蛋白（β-catenin）通路表达变化。方法：随机将SD孕鼠分为两组，一组孕鼠全程限食喂养，生产后通过减少喂养子鼠数量，建立子鼠CG-IUGR模型（CG-IUGR组）；另一组孕鼠正常喂养，生产的子鼠作为对照组。为排除性别因素的干扰，CG-IUGR组和对照组均选取雄性子鼠。CG-IUGR组和对照组在12周龄时进行葡萄糖耐量试验，并取肾周脂肪组织标本，通过苏木素-伊红染色观察脂肪结构，共聚焦显微镜下观察脂肪组织LRP6、β-catenin及胰岛素受体底物1（IRS-1）的免疫活性，蛋白质印迹法检测LRP6、β-catenin、IRS-1蛋白表达。结果：CG-IUGR组在60 min时血糖浓度及曲线下面积均大于对照组（均 P<0.05），表明CG-IUGR组葡萄糖耐量受损。CG-IUGR组脂肪细胞面积较对照组增大，脂肪组织中LRP6、β-catenin和IRS-1免疫活性较对照组降低（均 P<0.05）。     

子宫颈鳞状细胞癌中SCCA1和SCCA2的表达及意义

目的检测子宫颈鳞状细胞癌组织中SCCA(squamous cell carcinoma antigen)1和SCCA2 mRNA的表达,探讨其在子宫颈鳞状细胞癌临床诊断、疗效评价及预后观察中的作用。方法采用实时PCR的方法定量分析60例子宫颈鳞状细胞癌组织和30例正常子宫颈组织中SCCA1和SCCA2 mRNA的表达,并分析二者与临床病理的关系。结果 SCCA2 mRNA的表达在子宫颈鳞状细胞癌组织中较正常子宫颈组织中高,差异有统计学意义(P<0.001),而SCCA1 mRNA的表达则无统计学差异。SCCA2 mRNA的表达随着临床分期的增高而增高(P<0.001),SCCA2 mRNA的表达在有淋巴结转移组较无淋巴结转移组高(P<0.05),SCCA2 mRNA的表达与病理分级和年龄无关(P>0.05),SCCA1 mRNA的表达与年龄、病理分级、临床分期及淋巴结转移均无明显相关性(P>0.05)。结论 SCCA2 mRNA的表达可能为子宫颈鳞状细胞癌临床分期、淋巴结转移提供更为准确的判断信息。     

骨桥蛋白在口腔鳞状细胞癌中的定量表达及临床意义

目的:探讨骨桥蛋白(osteopontin,OPN)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)肿瘤组织中的表达及其临床意义.方法:EnVisionTM法检测正常口腔黏膜(n=12)和OSCC患者肿瘤组织(n=59)中OPN的表达,应用图像分析系统对染色结果进行定量分析,统计不同临床、病理指标下OSCC肿瘤组织OPN的表达情况.结果:OPN在OSCC肿瘤组织中的表达显著高于正常口腔黏膜(P＜0.05).OPN表达在OSCC不同临床分期、有无颈淋巴结转移间均有显著差异:临床Ⅲ～Ⅳ期(n=23)表达高于临床Ⅱ期(n=16),后者表达高于临床Ⅰ期(n=20)者(P均＜0.05);颈淋巴结转移患者(n=20)表达高于未转移者(n=39,P＜0.05).而其表达水平在OSCC高分化(n=38)和中低分化患者(n=21)间无显著差异.结论:OPN在OSCC中存在过度表达,其表达水平与肿瘤临床分期以及有无颈淋巴结转移存在一定的相关性(临床晚期患者、颈淋巴结转移患者表达程度较高),而与肿瘤分化程度无关.     

小檗碱通过激活Nrf2-HO-1/GPX4通路抑制小鼠海马神经元HT22细胞的铁死亡

目的探讨小檗碱对于Erastin诱导小鼠海马神经元HT22细胞的铁死亡的保护作用及其可能机制。方法以HT22小鼠海马神经元细胞为研究对象，分为对照组、Erastin模型组、Erastin+30 μmol/L BBR组、Erastin+60 μmol/L BBR组。采用CCK-8法、特异性Fe²⁺荧光探针、荧光染料（DAPI）检测和荧光探针（H2DCFH-DA）检测各实验组细胞的增殖情况、活性铁水平、细胞凋亡和活性氧（ROS）变化。RT-qPCR和Western blot分别检测各实验组细胞的Nrf2、HO-1、GPX4 mRNA和蛋白表达情况。以60 μmol BBR的最适浓度来进一步探究其作用机制，分为对照组、Erastin模型组、Erastin+60 μmol/L BBR组、Erastin+60 μmol/L BBR+2 μmol Nrf2抑制剂ML385组。通过使用荧光探针和Western blot检测Nrf2抑制剂（ML385）作用后的活性铁的水平、活性氧含量以及Nrf2、HO-1、GPX4蛋白的表达来验证小檗碱调节的Nrf2-HO-1/GPX4通路对Erastin处理的HT22细胞的保护作用。结果0.5 μmol/L Erastin作用于HT22细胞8 h，细胞存活率与对照组相比显著被抑制（P < 0.05）；同时细胞凋亡、ROS以及活性铁含量增加（P < 0.05）。与Erastin组比较，Erastin+30 μmol/L BBR组和Erastin+60 μmol/L BBR组的细胞存活率明显升高（P < 0.05），同时显著降低细胞凋亡、ROS以及活性铁含量（P < 0.05）。小檗碱增加HT22细胞中Nrf2、HO-1、GPX4基因及蛋白的表达量（P < 0.05）。加入Nrf2抑制剂ML385后，Nrf2-HO-1/GPX4通路被抑制，并且ROS以及活性铁含量升高（P < 0.05）。结论Erastin诱导HT22细胞发生铁死亡，小檗碱抑制Erastin诱导的铁死亡，可能机制是激活了Nrf2-HO-1/GPX4通路。     

食管鳞状细胞癌组织中MUC4和CerbB-2蛋白的表达

目的:探讨MUC4及CerbB-2的表达与食管鳞状细胞癌发生、发展及浸润转移的关系.方法:应用免疫组织化学SP法检测60例食管鳞状细胞癌组织、30例癌旁不典型增生组织和60例正常食管黏膜组织中MUC4及CerbB-2蛋白的表达.结果:食管鳞状细胞癌组织、癌旁不典型增生组织和正常食管黏膜组织中MUC4的阳性表达率分别为81.7%、56.7%和5.0%,CerbB-2的阳性表达率分别为73.3%、80.0%、36.7%,3组间MUC4与CerbB-2阳性表达率相比,差异均有统计学意义(P＜0.05).MUC4及CerbB-2的蛋白表达均与食管鳞状细胞癌的组织学分级、浸润深度及淋巴结转移密切相关(P＜0.05).结论:MUC4及CerbB-2在食管癌的浸润、转移及黏膜上皮癌变过程中起重要作用,2者联合检测可望成为食管鳞状细胞癌早期诊断和判断预后的分子指标之一.     

TGF-β1通过上调Shh信号诱导大鼠脑膜成纤维细胞向肌成纤维细胞转化

目的探讨TGF-β1在诱导脑膜成纤维细胞转化为肌成纤维细胞中对Shh信号的影响。方法新生24 h SD大鼠脑膜原代成纤维细胞经Ⅳ型胶原酶提取纯化。原代脑膜成纤维细胞随机分为3个组：分别为空白对照组、10 ng/mL TGF-β1处理组（TGF-β1组）、10 ng/mL TGF-β1联合20 μmol/L TGF-β1受体抑制剂SB-431542组(TGF-β1+SB组)。各组细胞在药物处理前均用无血清培养基培养24 h后换用加入了药物的DMEM/F12完全培养基处理72 h。采用CCK-8法检测各实验组成纤维细胞的增殖能力；细胞划痕实验检测各实验组成纤维细胞的迁移能力；免疫荧光法检测各实验组纤维连接蛋白（Fn）的表达；Western blotting检测各实验组Fn、α-平滑肌肌动蛋白（α-SMA）、Shh蛋白的表达；q-PCR检测各实验组细胞Shh mRNA的表达。结果TGF-β1可促进原代脑膜成纤维细胞的增殖（P < 0.05）；TGF-β1可促进原代脑膜成纤维细胞的迁移（P < 0.05）。TGF-β1可促进成纤维细胞向肌成纤维细胞的转化并增加Fn的分泌（P < 0.05）。TGF-β1处理后可上调Shh蛋白和mRNA的表达（P < 0.05），TGF-β1受体抑制剂SB-431542则可抑制TGF-β1引起的原代脑膜成纤维细胞向肌成纤维细胞的转化并减少纤维连接蛋白的分泌（P < 0.05）。结论TGF-β1可通过上调Sonic Hedgehog信号通路中Shh蛋白的表达，诱导脑膜成纤维细胞转化为肌成纤维细胞。     

早老蛋白1功能缺失性突变导致家族性阿尔茨海默病的研究进展

阿尔茨海默病（AD）是一种衰老相关的神经退行性疾病，主要病理特征是大脑中存在异常聚集的β淀粉样蛋白（Aβ）。AD可分为家族性和散发性，其中早老蛋白1（ PS1）是家族性AD最主要的风险基因， PS1突变占已知致家族性AD突变的80%以上。PS1是构成γ-分泌酶的催化亚基，后者负责加工Aβ前体蛋白（APP）生成Aβ。虽然新型 PS1突变日渐被报道，但其诱发家族性AD的分子机制仍无定论。由于90%的 PS1突变降低γ-分泌酶活性，学术界提出了 PS1功能缺失性突变假说，认为 PS1突变通过显性负效应导致PS1功能下降或缺失是诱发家族性AD的关键。近年，大量实验研究支持了该假说。首先， PS1功能缺失性突变通过干扰γ-分泌酶在APP上的切割位点促进长链Aβ生成，进而增加Aβ ₄₂/Aβ ₄₀比率；其次， PS1功能缺失性突变可破坏神经细胞内质网中的钙离子稳态以及通过阻断神经细胞自噬活性导致APP加工产物的异常聚集；再者， PS1功能缺失性突变可通过干扰神经元的内吞和转胞吞作用诱发神经元萎缩以及通过激活神经免疫细胞（星形胶质细胞和小胶质细胞）增强神经炎症；最后， PS1功能缺失性突变降低糖酵解和乳酸输出，破坏机体对神经元的能量供应。本文总结了 PS1功能缺失性突变诱发家族性AD的分子机制，并对今后潜在的研究方向进行了探讨。     

Occurrence and distribution of extended-spectrum β-lactamase in clinical Escherichia coli isolates at Ho Teaching Hospital in Ghana

ObjectiveThis study determined the occurrence and distribution of Extended Spectrum β-Lactamase (ESBL) genotypes of E. coli isolates in Ho Teaching Hospital, Ghana.DesignA cross-sectional study.SettingA single centre study was conducted at Ho Teaching Hospital of Ghana.ParticipantsPatients who visited Ho Teaching Hospital Laboratory with the request for culture and susceptibility testing.Main outcome measureEscherichia coli were isolated, and Extended-Spectrum β-Lactamase genes were detected.ResultsOf the 135 isolates, 56(41.5%,95% CI: 33.1% – 50.3%) were ESBL producers. More males, 14(58.3%), produced ESBL than females, 42(37.8%). The ESBL prevalence was highest among the elderly who were 80 years and above 3(100.0%), with the least prevalence among patients within 50–59 years and 0–9 years age bracket, representing 4(25.0%) and 3(27.3%), respectively. The total prevalence of ESBL was marginally higher among out-patients (41.8% 95% CI: 31.9% – 52.2%) compared to in-patients [40.5% 95% CI: 24.8% – 57.9]. BlaTEM-1 was the predominant ESBL genotype obtained from 83.9% (47/56) of the confirmed ESBL producing isolates, with the least being TOHO-1 4(7.1%). The co-existence of 2 different ESBL genes occurred in 19(33.9%) of the isolates. The single and quadruple carriage were 16(28.6%) and 3(5.4%), respectively. The highest co-existence of the ESBL genotypes was recorded for blaTEM-1 and blaCTXM-1 15(26.8%), followed by blaTEM-1, blaCTXM-1 and blaSHV-73 [12(21.4%)].ConclusionThe high prevalence of ESBL-producing E. coli isolates with multiple resistant gene carriage is a threat to healthcare in the study area.FundingThis research received no external funding.