LINC00649通过miR-424-5p/IGF1R轴调控内质网应激介导的宫颈癌细胞凋亡

目的探讨LINC00649/miR-424-5p/IGF1R对内质网应激(ERs)介导的宫颈癌(CC)细胞凋亡的影响。方法从GEO数据库中获取CC相关的数据,并分析差异表达的miRNAs。利用生物信息学数据库预测miR-424-5p的上、下游靶点,将LINC00649和IGF1R纳入研究,随后双荧光素酶实验进一步验证靶向关系。qRT-PCR检测LINC00649、miR-424-5p和IGF1R在CC组织和细胞中的表达水平。CCK-8和流式细胞术分别评估CC细胞增殖和凋亡变化。Western blot检测ERs相关蛋白GRP78、CHOP和Caspase-12的表达。结果与癌旁组织和H8细胞相比,LINC00649和IGF1R在CC组织和细胞中表达上调,而miR-424-5p下调(均P<0.05)。LINC00649的异常高表达与CC患者的预后不良有关,敲减LINC00649可通过促进ERs来抑制CC细胞活力,诱导细胞凋亡(均P<0.05)。LINC00649吸附miR-424-5p上调IGF1R的表达。miR-424-5p抑制剂或过表达IGF1R均可部分逆转敲减LINC00649对CC细胞的影响(均P<0.05)。结论 LINC00649能够通过miR-424-5p/IGF1R抑制CC细胞的ERs过程进而减少细胞凋亡,提高细胞活力。     

新型降脂药人前蛋白转化酶枯草溶菌素 9抑制剂与动脉粥样硬化性心血管疾病、静脉血栓栓塞疾病关系的研究进展

随着生活方式的改变，心脑血管疾病逐渐成为全人类死亡的“头号杀手”高于恶性肿瘤、糖尿病等。血脂的升高尤其是低密度脂蛋白胆固醇( LDL-C)的升高与动脉粥样硬化性心血管疾病( ASCVD)的发，生、发展息息相关。现有的专家共识、南提出他汀类药物是降低 LDL-C的一线用药，但仍可能会发生复发性缺血事件。人前蛋白转化酶枯草溶菌素 9(PCSK9)抑制指剂作为一类新型降脂药，有显著降低血 LDL-C水平，同时又可降低脂蛋白( a)的水平，又与 ASCVD与静脉血栓栓塞疾病( VTE)的发病可能相关。本文将系统地阐述新型降脂药 PCSK9抑制剂与 ASCVD、VTE的关系研究进展。     

Immunity of two novel hepatitis C virus polyepitope vaccines

Hepatitis C virus (HCV) infection remains a serious public health burden around the world. So far there is no effective vaccine against this virus. Neutralizing antibody (NAb) responses to the epitopes within HCV E1 and E2 proteins are related to the resolution of hepatitis C infection. E. coli heat-labile enterotoxin B subunit (LTB) has been described as potent immunity adjuvants. In this study, we constructed recombinant pET vectors: pET-R9-Bp (B cell polyepitopes) expressing 7 epitopes from HCV E1 and E2 proteins including R9 (E2_384-411aa)-Bp (E1_313-327aa-E2_396-424aa-E2_436-447aa-E2_523-540aa-E2_610-627aa-E2_631-648aa) and pET-LTB-R9-Bp expressing LTB adjuvant in combination with R9-Bp. Recombinant proteins R9-Bp and LTB-R9-Bp were expressed successfully in E. coli and purified by the Ni-NTA column. Both R9-Bp and LTB-R9-Bp in BALB/c mice induced robust humoral immune response in the context of intraperitoneal or intramuscular immunization but not oral immunization. Intraperitoneal administration of LTB-R9-Bp induced a higher antibody titer (peak titer: 1:341000) than that of R9-Bp (peak titer: 1:85000) after the second boost (P = 0.0036 or 0.0002). However, comparable antibody peak titers were elicited for both R9-Bp and LTB-R9-Bp in intramuscular immunization albeit with significant difference (P = 0.0032) a week after the second boost. In addition, both R9-Bp and LTB-R9-Bp induced the secretion of cytokines including IFN-γ and IL-4 at similar levels. anti-sera induced by both R9-Bp and LTB-R9-Bp recognized native HCV E1 and E2 proteins. Moreover, these HCV-specific antisera inhibited significantly the entry of HCV (P < 0.0001). Taken together, these findings showed that E. coli-based both R9-Bp and LTB-R9-Bp could become promising HCV vaccines.     

Frequency-specific effects of low-intensity rTMS can persist for up to 2 weeks post-stimulation: A longitudinal rs-fMRI/MRS study in rats

BackgroundEvidence suggests that repetitive transcranial magnetic stimulation (rTMS), a non-invasive neuromodulation technique, alters resting brain activity. Despite anecdotal evidence that rTMS effects wear off, there are no reports of longitudinal studies, even in humans, mapping the therapeutic duration of rTMS effects.ObjectiveHere, we investigated the longitudinal effects of repeated low-intensity rTMS (LI-rTMS) on healthy rodent resting-state networks (RSNs) using resting-state functional MRI (rs-fMRI) and on sensorimotor cortical neurometabolite levels using proton magnetic resonance spectroscopy (MRS).MethodsSprague-Dawley rats received 10 min LI-rTMS daily for 15 days (10 Hz or 1 Hz stimulation, n = 9 per group). MRI data were acquired at baseline, after seven days and after 14 days of daily stimulation and at two more timepoints up to three weeks post-cessation of daily stimulation.Results10 Hz stimulation increased RSN connectivity and GABA, glutamine, and glutamate levels. 1 Hz stimulation had opposite but subtler effects, resulting in decreased RSN connectivity and glutamine levels. The induced changes decreased to baseline levels within seven days following stimulation cessation in the 10 Hz group but were sustained for at least 14 days in the 1 Hz group.ConclusionOverall, our study provides evidence of long-term frequency-specific effects of LI-rTMS. Additionally, the transient connectivity changes following 10 Hz stimulation suggest that current treatment protocols involving this frequency may require ongoing “top-up” stimulation sessions to maintain therapeutic effects.     

Platelets are important for the development of immune tolerance: Possible involvement of TGF‐β in the mechanism

Platelets have diverse roles in immune processes in addition to their key functions in haemostasis and thrombosis. Some studies imply that platelets may be possibly related to the immune tolerance induction. However, the role of platelets in the development of immune tolerance is not fully understood. The purpose of this study was to investigate the role of platelets in the development of regulatory mechanisms responsible for cutaneous inflammation using a mouse model of low zone tolerance (LZT). Mice were treated with 2,4,6‐trinitro‐1‐chlorobenzene (TNCB) 8 times every other day for tolerance induction with administration of anti‐platelet antibody or control antibody during the tolerance induction phase every 3 days. After the treatment for the tolerance induction, mice were sensitized and then challenged with TNCB. The contact hypersensitivity (CHS) was significantly decreased at 24 hours after challenge in the mice with LZT than in those without LZT. Platelet depletion via administration of anti‐platelet antibody reversed the inhibition of CHS and reduced the frequency of Foxp3⁺ Tregs in the inflamed skin and draining lymph nodes in mice with LZT. In addition, repeated low‐dose skin exposure resulted in elevated plasma levels of transforming growth factor (TGF)‐β1. Interestingly, platelet depletion reduced plasma TGF‐β1 levels of mice with LZT. Furthermore, the CHS response was reduced by administration of recombinant TGF‐β1 during platelet depletion in mice with LZT. Administration of anti‐TGF‐β antibody reversed the inhibition of the CHS responses. These results suggest that platelets are involved in the induction of immune tolerance via the release of TGF‐β1.     

A new cause of developmental and epileptic encephalopathy with continuous spike-and-wave during sleep: CDKL5 disorder

The CDKL5 disorder is characterized by early onset epilepsy, stereotypical hand movement, absent speech and severe hypotonia. Herein, we report epileptic encephalopathy with continuous spike-and-wave during sleep (CSWS) in apatient with CDKL5 disorder. She admitted with complaints of frequently recurring generalized tonic and myoclonic seizures. The diagnoses were confirmed by de novo CDKL5 mutation, c.197_198delCT (p.L67QfsX23). Interictal EEG revealed generalized spike and slow-wave activity, occurring intermittently in wakefulness but present for at least 85% of non-REM sleep, consistent with the diagnosis of CSWS. To our knowledge, this is the first report of CSWS associated with CDKL5 disorder.     

改良跗骨窦入路治疗 SandersⅡ～Ⅲ 型跟骨骨折疗效分析

目的探讨改良跗骨窦入路与传统跗骨窦入路治疗 SandersⅡ～Ⅲ 型跟骨骨折的近期临床疗效。方法选取 2015 年 1 月—2017 年 8 月符合选择标准的 53 例 SandersⅡ～Ⅲ 型跟骨骨折患者，采用随机数字表法将患者分为观察组（21 例，采用从腓骨长短肌腱下方暴露距下关节后对骨折复位的改良跗骨窦入路）和对照组（32 例，采用传统跗骨窦入路）。两组患者性别、年龄、侧别、致伤原因、骨折分型、受伤至手术时间以及术前 Böhler 角、Gissane 角、疼痛视觉模拟评分（VAS）等一般资料比较差异均无统计学意义（P>0.05），具有可比性。记录并比较两组患者手术时间、术后引流量、手术相关并发症，术后 Böhler 角、Gissane 角及术后角度改善值；并使用 VAS 评分、美国矫形足踝协会（AOFAS）踝与后足评分及简明健康调查量表（SF-36 量表）评价疗效。 结果53 例患者均顺利完成手术，无血管神经损伤、围手术期死亡等严重并发症。两组患者手术时间及术后引流量比较差异无统计学意义（P>0.05）。两组患者均获随访，随访时间 12～36 个月，平均 17 个月。术后未发生骨折移位、内固定失败、骨折畸形愈合等；所有患者均未行二期关节融合手术。两组骨折愈合时间比较差异无统计学意义（t=0.30，P=0.77）。两组术后 2 d Böhler 角、Gissane 角均较术前显著改善（P<0.05）；但观察组术后 2 d Böhler 角、Gissane 角及改善值与对照组比较差异均无统计学意义（P>0.05）。两组术后 24 h 及 1 年 VAS 评分均较术前显著改善（P<0.05）；术后 24 h 及 1 年两组 VAS 评分比较差异均无统计学意义（P>0.05）。两组术后 1 年 AOFAS 评分比较差异无统计学意义（t=1.46，P=0.15）；但观察组术后 1 年 SF-36 量表评分显著高于对照组（t=2.08，P=0.04）。末次随访时，观察组 2 例、对照组 8 例患者出现距下关节僵硬或疼痛，两组发生率比较差异无统计学意义（χ²=1.98，P=0.16）。 结论采用改良跗骨窦入路治疗 SandersⅡ～Ⅲ 型跟骨骨折，具有创伤小、直视下复位清晰、复位固定可靠、切口并发症少等优点。