Distribution of Oxytocin Hypothamic-Picuitary-Ovary Axis in Guangxi Local Buffalo

To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochemistry SuperPicTureTM two step method was used to detect the distribution of OT in hypothalamatic-pituitary-ovary axis of five femal Guangxi local buffalo. The test results could provide morphology according to study the OT's synthesis and mechanism of action,and could play reference and directions part in breeding Guangxi local buffalo. The test results display:oxytocin immuno reactive (OT-IR) neuronsw eremainly distributed arcuate nucleus,supraoptic nucleus and paraventricular nucleus,and OT-IR neurons was also found in ventromedial nucleus,ventrolateralis nucleus,suprachiasmaticus nucleus,dorsomedial nucleus,mamillary body,anterior hypothalamic nucleus and so on. The OT immunoactive production was found in pituitary and few OT-IR nerve fibers extended to post pituitary from hypophyseal stalk and medium eminence. In ovaries,OT immunoactive productions were only distributed in germinal epithelium cells,granulosa cells and lutein cells. The OT was first discovered in singulorum link of hypothalamatic-pituitary-ovary axis of Guangxi local buffalo. The OT immunoactive neurons were first discovered in every main nucleus of Guangxi local buffalo hypothalamus,especially distributed in arcuate nucleus,supraoptic nucleus and paraventricular nucleus.

β-CTX、t-P1NP及N-MID-OT在绝经后女性骨质疏松性骨折风险评估中的应用

目的探讨β胶原特殊序列(β-crosslaps,β-CTX)、总1型胶原氨基端延长肽(t-P1NP)及N端中段骨钙素(N-MID-OT)在绝经后女性骨质疏松性骨折风险评估中的应用价值。方法回顾性分析2020年2月至2022年12月解放军总医院第六医学中心收治的102例绝经后骨质疏松(postmenopausal osteoporosis,PMOP)患者的临床资料,将其作为研究组,并根据是否发生骨折将其分为PMOP骨折组(39例)与PMOP未骨折组(63例),另选100名健康体检者作为对照组。记录所有研究对象的β-CTX、t-P1NP及N-MID-OT水平并进行比较,用ROC曲线评价β-CTX、t-P1NP及N-MID-OT在绝经后女性骨质疏松性骨折风险评估中的应用价值。结果研究组的平均体重、体质量指数、股骨颈骨密度(bone mineral density,BMD)、左髋总和BMD及L 1~4总和BMD均明显低于对照组(P<0.05);而两组研究对象的平均年龄、平均身高及绝经年龄等比较差异均无统计学意义(P>0.05)。研究组的β-CTX、t-P1NP及N-MID-OT均明显高于对照组(t值依次为12.688、37.430、26.599,P<0.05)。β-CTX+t-P1NP+N-MID-OT联合检测用于预测PMOP的AUC值为0.978,敏感度为98.04%,特异度为97.00%,表明β-CTX+t-P1NP+N-MID-OT三指标联合检测用于预测PMOP的效能更高。PMOP骨折组的β-CTX、t-P1NP及N-MID-OT均明显高于PMOP未骨折组(t值依次为6.078、16.363、12.227,P<0.05)。β-CTX+t-P1NP+N-MID-OT联合检测用于评估PMOP骨折风险的AUC值为0.939,敏感度为94.87%,特异度为95.24%,表明β-CTX+t-P1NP+N-MID-OT联合检测用于评估PMOP骨折风险的效能更高。结论PMOP患者β-CTX、t-P1NP及N-MID-OT水平均明显升高,而PMOP骨折患者β-CTX、t-P1NP及N-MID-OT水平升高更为明显,且β-CTX+t-P1NP+N-MID-OT联合检测可显著提高对PMOP预测及骨折风险评估效能,值得借鉴。

阿仑磷酸钠辅助PVP治疗对老年骨质疏松性压缩性骨折BALP、PTH及N-MID-OT水平的影响

目的分析阿仑磷酸钠辅助经皮椎体成形术(PVP)治疗对老年骨质疏松性压缩性骨折骨碱性磷酸酶(BALP)、甲状旁腺激素(PTH)及N端中段骨钙素(N-MID-OT)水平的影响。方法选取2019年1月至2022年2月唐山市丰润区人民医院收治的老年骨质疏松性压缩性骨折患者107例,按照治疗方式不同分为观察组和对照组,对照组采取PVP治疗(n=52),观察组采取阿仑磷酸钠辅助PVP治疗(n=55)。对比两组Oswestry功能障碍指数问卷表(ODI)、视觉模拟量表(VAS)、骨代谢指标(BALP、PTH、N-MID-OT水平)、不良反应及再骨折发生率。结果两组术后2个月、6个月、12个月ODI评分均下降,且观察组术后各时间段ODI评分均低于对照组,差异有统计学意义(P<0.05)。两组术后2个月、6个月、12个月VAS评分均下降,且观察组术后各时间段VAS评分均低于对照组,差异有统计学意义(P<0.05)。两组术后BALP、PTH及N-MID-OT水平均下降,且观察组术后BALP、PTH及N-MID-OT水平均低于对照组,差异有统计学意义(P<0.05)。两组不良反应总发生率对比,差异无统计学意义(P>0.05),但观察组再骨折发生率(7.28%)明显低于对照组(25.00%),差异有统计学意义(P<0.05)。结论阿仑磷酸钠辅助PVP治疗可有效改善老年骨质疏松性压缩性骨折患者椎体功能,减轻其疼痛程度,改善患者体内BALP、PTH及N-MID-OT水平,促进其术后恢复。

不明原因复发性流产患者血清lncRNA-KCNQ1OT1、miR-29a-3p及SOCS3mRNA水平的表达及其相关性

复发性流产是指与同一性伴侣在妊娠28周之前,连续2次及以上发生自然流产的情况,其病因复杂,至今仍有50%以上的复发性流产病因尚未明确,即不明原因复发性流产(URSA)^([1-2])。有研究表明URSA患者子宫蜕膜存在明显的炎症微环境,可以引起性激素紊乱,与URSA的发生及发展密切相关^([3])。细胞因子信号传导抑制因子3 (SOCS3)是一个具有多项调节功能的蛋白质分子,可以调节机体免疫炎症反应,能够促进滋养细胞和T细胞增殖分化,从而影响着复发性流产的发生和发展^([4-5])。

Oxytocin decreases colonic motility of cold water stressed rats via oxytocin receptors

AIM: To investigate whether cold water intake into the stomach affects colonic motility and the involvement of the oxytocin-oxytocin receptor pathway in rats. METHODS: Female Sprague Dawley rats were used and some of them were ovariectomized. The rats were subjected to gastric instillation with cold (0-4 degrees C, cold group) or room temperature (20-25 degrees C, control group) saline for 14 consecutive days. Colon transit was determined with a bead inserted into the colon. Colonic longitudinal muscle strips were prepared to investigate the response to oxytocin in vitro. Plasma concentration of oxytocin was detected by ELISA. Oxytocin receptor expression was investigated by Western blot analysis. Immunohistochemistry was used to locate oxytocin receptors. RESULTS: Colon transit was slower in the cold group than in the control group (P < 0.05). Colonic smooth muscle contractile response to oxytocin decreased, and the inhibitory effect of oxytocin on muscle contractility was enhanced by cold water intake (0.69 +/- 0.08 vs 0.88 +/- 0.16, P < 0.05). Atosiban and tetrodotoxin inhibited the effect of oxytocin on colonic motility. Oxytocin receptors were located in the myenteric plexus, and their expression was up-regulated in the cold group (P < 0.05). Cold water intake increased blood concentration of oxytocin, but this effect was attenuated in ovariectomized rats (286.99 +/- 83.72 pg/mL vs 100.56 +/- 92.71 pg/mL, P < 0.05). However, in ovariectomized rats, estradiol treatment increased blood oxytocin, and the response of colonic muscle strips to oxytocin was attenuated. CONCLUSION: Cold water intake inhibits colonic motility partially through oxytocin-oxytocin receptor signaling in the myenteric nervous system pathway, which is estrogen dependent. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.

Oxytocin but Not Testosterone Modulates Behavioral Patterns in Autism Spectrum Disorders

Autism spectrum disorder (ASD) is a neurodevelopmental disorder of unknown etiology. Social deficits represent one of the core symptoms of the diagnosis. The aim was to reveal possible correlations among peripheral levels of oxytocin and testosterone with behavioral and symptom characteristics in patients with ASD. 8 children with ASD were recruited and underwent psychological profiling. Blood oxytocin and testosterone levels were analyzed using ELISA method. Oxytocin levels positively correlated with Adaptation to change category of CARS-2 (P = 0.008, R = 0.848) and Vineland-II maladaptive behavior scores (P = 0.004, R = 0.884). No significant correlations were found among testosterone levels and behavioral parameters. Higher oxytocin levels were connected with more severe adaptive behavior in ASD patients. Increased oxytocin levels in children with more severe phenotype could be a result of compensatory mechanism of impaired oxytocin signaling. Oxytocin seems to employ distinct mechanisms in regulating social behavior in autism and healthy population.

The Role of Ankaferd Blood Stopper and Oxytocin as Potential Therapeutic Agents in Endometriosis:A Rat Model

To evaluate the potential effect of Ankaferd Blood Stopper(ABS)and oxytocin(OT)in an experimental endometriosis model,18 female Sprague Dawley rats were used in this study.The animals were divided randomly into three groups after surgical induction of endometriosis:group 1:control group(isotonic NaCl,1 mL/kg/day,intramuscular,n=6);group 2:OT group(OT,80 U/kg/day,intramuscular,n=6);group 3:ABS group(ABS,1.5 mL/kg/day,intraperitoneal,n=6).Each group was treated for four weeks(two times per week).Volumes of endometriotic explants were measured in biopsy samples for histopathological analysis.Vascular endothelial growth factor(VEGF),monocyte chemotactic protein-1(MCP-1),and tumour necrosis factor(TNF-α)levels were measured in plasma and peritoneal fluid.Endometriotic explant volumes were significantly decreased after OT administration(P<0.0001).The epithelial score was significantly decreased in both treatment groups compared to the control group(P<0.05).TUNEL immunohistochemistry showed more apoptotic changes in the endometriosis foci(gland epithelium and surrounding tissue)in the OT group than in the control group(P<0.05).The levels of VEGF,MCP-1,and TNF-αwere significantly reduced in the OT group(P<0.05),whereas no significant changes in protein levels were found in the ABS-applied group.The results indicate that OT has greater potential as a therapeutic agent in experimentally induced peritoneal endometriosis,where ABS,which is a VEGF modulator,appears to act through different mechanisms to show its palliative effects on a rat model of peritoneal endometriosis.

The Correlation between Synthetic Oxytocin Given during Labor and Women’s Personality Traits According to the SSP (Swedish University Scales of Personality) Survey: A Pilot Study

Background: In modern obstetric care, oxytocin is one of the most frequently used drugs, and the possible mental impact this drug has on women is very little studied. The objective of this study is to investigate whether women augmented with oxytocin during labor will rate their personality profile differently after childbirth than non-stimulated women. Methods: Prospective cohort study was performed at Women’s Clinic, Soder hospital, Stockholm.76 women received the SSP (Swedish University Scales of Personality) questionnaire to fill in during their stay in the post-maternity ward after labor. Information about the use of oxytocin was retrieved from the women’s medical records. Primary outcome: Differences in the SSP scores in the group augmented with synthetic oxytocin during labor compared with the non-augmented group. Results: Women with and without oxytocin estimates on the SSP subscale form differed regarding personality traits described as “lack of assertiveness” (p = 0.04), which means “lack of ability to speak up and to be self-assertive in social situations”. The result also showed that women that had a long time of augmentation with oxytocin (>5 h) scored higher for “social desirability” (p = 0.004), which was defined as being “socially adapted,” “friendly,” and “helpful”. A difference in “psychological anxiety” (p = 0.04) and “social desirability” (p = 0.004) was found among women who had oxytocin in a dose of at least 200 ImU/h for ≥1 hour. This group also had a lower rate of “mental anxiety” than those who received lower oxytocin doses. Conclusion: Synthetic oxytocin given during labor may affect the woman mentally. The total time and volume of given oxytocin seem to be essential factors when discussing augmentation’s maternal psychological response. We conclude that prolonged and extended use of synthetic oxytocin during labor should be avoided if possible.

Effects of two different protocols of oxytocin infusion for labor induction on obstetric outcomes: A cohort study

Objective: Concerns remain about the safety and efficacy of high dose and low dose protocols of oxytocin for labor induction. We have compared 2 regimens of oxytocin induction (low-dose vs high dose) on perinatal outcomes over a 1-year period. Study Design: Included were all women undergoing induction of labor at term with live singleton gestations. Cesarean delivery (CD) and a composite adverse neonatal outcome (5-min Apgar score < 7, umbilical artery pH < 7.10, or need for admission to NICU) were assessed using logistic regression analysis. Admission-to-delivery intervals was compared between the two groups by log-rank test. Results: A total of 544 women fulfilled the study criteria. The two groups were comparable for demographic and obstetric variables. There was no significant association between oxytocin regimen and rates of CD (P = 0.77) or adverse neonatal outcome (P = 0.99) even after controlling for confounders. The admission-to-delivery interval was significantly shorter for the high-dose group than for the low-dose group (median interval = 11.7 vs 14.3 hours, respectively, P = 0.026). Conclusion: Use of a high-dose protocol of oxytocin administration for induction of labor at term is associated with similar rates of cesarean section and adverse neonatal outcome as a low-dose protocol, but with an average of 2.5 hours shorter duration of labor.

脑卒中患者采取数字OT训练系统与作业疗法干预对上肢功能和生活自理能力的影响分析

目的:探讨数字OT训练系统与作业疗法在脑卒中疾病中联合应用,对生活自理能力(ADL)评分和上肢功能的影响。方法:选取2020年5月—2021年5月天津市宝坻区人民医院收治的68例脑卒中患者作为研究对象,按照随机数表法分为对照组和观察组,每组各34例。对照组采取作业疗法,观察组采取作业疗法联合数字OT训练系统疗法。对比两组患者康复效果、训练配合度、ADL评分、肢体运动功能(Fugl-Meyer)评分、神经功能(NIHSS)评分、认知功能(MoCA)评分、健康生活质量(SF-36)评分等指标。结果:观察组患者临床康复效果高于对照组,差异有统计学意义(χ^(2)=5.100,P>0.05)。观察组患者训练配合度明显高于对照组,差异有统计学意义(χ^(2)=4.220,P>0.05)。干预后,观察组患者ADL评分、上肢功能评分高于对照组,差异有统计学意义(t=7.035、3.475,P<0.05)。干预后,观察组患者NIHSS评分低于对照组、MoCA评分高于对照组,差异有统计学意义(t=15.561、2.552,P<0.05)。干预后,观察组患者生活质量评分显著高于对照组,差异有统计学意义(t=8.457、7.053、4.750、3.534、7.755、3.782、7.559、4.507,P<0.05)。结论:脑卒中患者采取数字OT训练系统与作业疗法联合干预的效果明显,可改善其认知、神经功能,经合理训练后其上肢功能恢复明显,提升患者日常ADL、生活质量及训练配合度。

lncRNA KCNQ1OT1靶向调控miR-132-5p对帕金森病细胞损伤的保护机制

目的探究长链非编码RNA KCNQ1OT1(lncRNA KCNQ1OT1)靶向调控微小RNA-132-5p(miR-132-5p)对帕金森病细胞损伤的保护机制。方法SH-SY5Y细胞通过1-甲基-4-苯基吡啶阳离子(MMP+)诱导建立帕金森病细胞模型,检测SH-SY5Y细胞中lncRNA KCNQ1OT1、miR-132-5p表达、活性氧(ROS)、超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平、凋亡率及Bcl-2、Bax蛋白表达,验证lncRNA KCNQ1OT1、miR-132-5p的调控关系。结果与Con组相比,MMP+组lncRNA KCNQ1OT1、miR-132-5p表达量较高(P<0.05)。低表达lncRNA KCNQ1OT1、干扰miR-132-5p表达均可减轻MMP+诱导的SH-SY5Y细胞氧化应激及炎性损伤,抑制细胞死亡,lncRNA KCNQ1OT1靶向正调控miR-132-5p表达,miR-132-5p过表达逆转了lncRNA KCNQ1OT1低表达对MMP+诱导SK-N-SH细胞氧化应激、炎症损伤及凋亡。结论lncRNA KCNQ1OT1通过靶向抑制miR-132-5p表达减轻了MMP+诱导SK-N-SH细胞氧化应激、炎症损伤,抑制了细胞凋亡。

OTS自组装多层膜的制备和能量耗散的研究

采用自组装技术在石英表面制备了十八烷基三氯硅烷(OTS)分子膜,并通过接触角、红外光谱(FT-IR)和原子力显微镜(AFM)研究了膜层形貌特征和膜层表面能量耗散随温度的变化情况。结果表明:OTS在石英上以“岛”状团簇的结构均匀分布,反应时间越长,“岛”状团簇和吸附的OTS分子数量也越多,表面粗糙度随之增加。接触角在反应时间达到30min后稳定在110.39°左右,此时反应趋于饱和。膜层表面耗散值在60~80℃内最低,当温度低于61℃时,5mmol/L溶度下制备的分子膜表面最平坦稳定,膜层耗散值也最低,温度高于61℃时,表面出现熔化现象,耗散曲线波动较大,考虑是水解的OTS产物和表面杂质的存在影响了测试结果。

化工行业数字化转型背景下IT和OT技术融合路径研究

随着物联网、数字孪生技术的日臻完善,以及元宇宙概念的兴起,制造范式持续进化,实体与虚拟世界的融合应用场景日益丰富。在此背景下IT与OT的深度融合不仅是顺应“双碳”减排目标及化工产业升级的必然趋势,更是实现化工行业全面数字化转型的必然路径。本文深挖化工行业特性,系统性总结了化工行业数字化转型的核心要素,并针对IT与OT的融合路径进行了详尽剖析与设计,构建了一个高效协同的技术框架。此外,本研究基于工业互联网平台,将上述设计理念转化为实际应用,有效推动了化工企业的数字化转型升级,显著提升了企业生产效率与产品质量,为化工行业数字化转型树立了实践典范。

工业互联网环境下IT/OT融合的安全防御技术研究

传统IT/OT融合的安全防御技术只能储存IT/OT融合数据,不能进行加密,导致数据出现泄露、完整性受损以及真实性问题。为此,文章研究工业互联网环境下IT/OT融合安全防御技术。首先,文章通过风险评估、漏洞扫描、网络监控和日志分析等方法,寻找并确定安全缺口,构建安全架构关联体系;其次,建立防火墙体系,对网络进行主动优化和保护,确保不同区域的安全隔离;最后,采用加密技术对IT/OT融合数据进行加密处理,确保数据来源的真实可信性。实验结果表明,文章设计的安全防御技术攻击成功率最高仅为2%,具备更好的防御性能。

数值模式降水预报OTS订正法的实现技术

本文以ECMWF模式的逐12 h累计降水预报的订正试验为例,以福建、江西、浙江和上海为研究区域,详细介绍了福建省气象台在实现最优TS评分(Optimal Threat Score,OTS)订正法时引用或首创的三项技术,并与同类技术进行对比。结果表明:(1)用3年期准对称滑动窗口取样法采集训练样本计算OTS订正阈值,其效果优于取之前3年的当季样本的取样方法。(2)OTS订正阈值F1(即消空阈值)的大小与最高2 m温度密切相关,随着温度升高,消空阈值先变大后减小。相比不分组的方案,基于最高2 m温度预报分组计算消空阈值的方案能同时降低小雨的空报率和漏报率,使ETS评分提高5.0%~8.2%。(3)先用反距离加权插值法对模式降水预报进行插值,再应用OTS订正法对降水预报进行订正,其效果优于先用最邻近点插值法对模式降水预报进行插值,再应用OTS订正法对降水预报进行订正。

Growth hormone promotes the reconstruction of injured axons in the hypothalamo-neurohypophyseal system

Previous studies have shown that growth hormone can regulate hypothalamic energy metabolism, stress, and hormone release. Therefore, growth hormone has great potential for treating hypothalamic injury. In this study, we established a specific hypothalamic axon injury model by inducing hypothalamic pituitary stalk electric lesions in male mice. We then treated mice by intraperitoneal administration of growth hormone. Our results showed that growth hormone increased the expression of insulin-like growth factor 1 and its receptors, and promoted the survival of hypothalamic neurons, axonal regeneration, and vascular reconstruction from the median eminence through the posterior pituitary. Altogether, this alleviated hypothalamic injury-caused central diabetes insipidus and anxiety. These results suggest that growth hormone can promote axonal reconstruction after hypothalamic injury by regulating the growth hormone-insulin-like growth factor 1 axis.

OT和IT融合场景下网络安全威胁监测分析系统设计

在当今数字化时代,运营技术与信息技术的融合为企业带来了前所未有的效率和创新。企业在享受数字化转型带来的便利与效率提升的同时,也面临着日益复杂的网络安全威胁。工业控制系统的互联互通,使得潜在的安全漏洞和攻击面不断扩大,使得工业控制系统暴露在更广泛的网络安全威胁之下。将深入探讨运营技术与信息技术融合场景下的网络安全威胁监测,分析潜在风险,并提出有效的监测策略,以确保关键基础设施的安全与稳定。

血浆长链非编码RNASOX2-OT在老年慢性阻塞性肺疾病急性加重相关性肺动脉高压患者中的临床意义

目的检测血浆长链非编码RNA SOX2-OT在老年慢性阻塞性肺疾病急性加重(AECOPD)相关性肺动脉高压(PH)患者血浆中的表达水平,探讨其潜在的临床应用价值。方法前瞻性队列研究,纳入120例确诊为AECOPD的老年患者,根据肺动脉收缩压(PASP)分为2组:A组(AECOPD组,60例)和B组(AECOPD相关PH组,60例)。在入院时及入院后10 d,评估并比较2组血浆长链非编码RNA SOX2-OT水平、肺动脉收缩压(PASP)、炎症标志物血浆IL-6和IL-8水平、心脏生物标志物血浆NT-ProBNP水平、动脉血气指标值、肺功能指标值、6分钟步行距离测试结果。采用描述性统计分析2组的基线特征,使用t检验或非参数检验比较2组间的差异。并进行相关性分析以评估SOX2-OT与其他指标的关系。结果在入院时及入院后10 d,2组PASP、血浆长链非编码RNA SOX2-OT水平、血浆IL-6水平、血浆IL-8水平、血浆NT-ProBNP水平、动脉血气指标(pH值、PaO_(2)、PaCO_(2))值、肺功能指标(FEV 1%、DLCO)值、6分钟步行距离差异均有统计学意义(均P<0.05)。对B组患者从入院到入院后10 d的指标变化进行比较,发现PASP值、血浆SOX2-OT水平、炎症标志物水平、心脏生物标志物水平、动脉血气指标值、肺功能指标值、6分钟步行距离均显示出时间效应(均P<0.05)。在A组和B组中,血浆长链非编码RNA SOX2-OT水平与PASP值、血浆IL-6水平、血浆IL-8水平、血浆NT-ProBNP水平、PaCO_(2)值显示出显著的正相关性(均P<0.05),而与pH值、PaO_(2)值、FEV 1%值、DLCO值、6分钟步行距离呈现出显著的负相关性(均P<0.05)。结论合并PH的AECOPD老年患者血浆长链非编码RNA SOX2-OT水平明显高于不合并PH的AECOPD老年患者;AECOPD相关PH患者经过控制感染、纠正呼吸衰竭等治疗后,随着肺动脉压力下降,血浆长链非编码RNA SOX2-OT水平明显下降。

LncRNA SOX2OT靶向SIRT1/自噬通路增强胆管癌细胞5-FU耐药

目的探讨LncRNASOX2OT靶向SIRT1/自噬通路调节胆管癌细胞5-FU耐药的机制。方法将未用5-FU处理HCCC-9810细胞和不同浓度(50、100、150、200μg/mL)5-FU处理的HCCC-9810细胞分为对照组和模型组,qRT-PCR检测LncRNA SOX2OT、SIRT1 mRNA表达水平,Western blot检测SIRT1、Beclin1、LC3和p62蛋白表达。pcDNA3.1-SOX2OT和pcDNA3.1-NC转染HCCC-9810/5-FU耐药细胞,CCK-8检测细胞耐药性,划痕实验检测细胞迁移能力,qRT-PCR检测LncRNASOX2OT、SIRT1 mRNA表达水平,Western blot检测SIRT1、Beclin1、LC3和p62表达。在以上基础上做了Rescue实验,将OV-SOX2OT(2μg/孔)和si-NC(75pmol/孔)、OV-SOX2OT(2μg/孔)和si-SIRT1(75pmol/孔)分别共转染HCCC-9810/5-FU耐药细胞,分组为OV-SOX2OT+si-NC组和OV-SOX2OT+si-SIRT1组,以证明LncRNASOX2OT通过SIRT1影响自噬,并由此影响胆管癌细胞对5-FU的耐药性。RNAPulldown验证SOX2OT与SIRT1的靶向结合关系。结果不同浓度的5-FU均能抑制HCCC-9810细胞增殖(P<0.05)。与对照组相比,模型组SIRT1、Beclin1(P<0.001)和p62(P<0.01)蛋白表达、LC3Ⅱ/LC3Ⅰ比值(P<0.001)、SIRT1和LncRNA SOX2OT mRNA水平(P<0.05)均明显增加。与OV-NC组相比,OV-SOX2OT组细胞迁移能力、SIRT1、Beclin1(P<0.001)和p62(P<0.05)蛋白表达、LC3Ⅱ/LC3Ⅰ比值(P<0.001)、SIRT1和LncRNASOX2OTmRNA(P<0.05)水平均明显增加。沉默SIRT1表达导致LncRNASOX2OT过表达的HCCC-9810细胞对5-FU的耐药性显著降低。与OV-SOX2OT+si-NC组相比,OV-SOX2OT+si-SIRT1组SIRT1(P<0.001)、p62和Beclin1(P<0.01)蛋白表达、LC3Ⅱ/LC3Ⅰ比值(P<0.01)、SIRT1 mRNA(P<0.05)水平均明显降低。RNA Pulldown检测结果显示SOX2OT能够直接与SIRT1结合。结论LncRNA SOX2OT通过上调SIRT1表达促进自噬来增强胆管癌HCCC-9810细胞5-FU耐药性。

Role of oncogenic long noncoding RNA KCNQ1OT1 in colon cancer

The role of lncRNA KCNQ1 opposite strand/antisense transcript 1(KCNQ1OT1)in colon cancer involves various tumorigenic processes and has been studed widely.However,the mechanism by which it promotes colon cancer remains unclear.Retrovirnl vector pSEB61 was retroftted in established HCT116 siKCN and SW480-siKCN cells to silence KCNQ1 OT1.Cellular proliferation was measured using CCK8 assay,and flow cytometry(FCM)detected cell cydle changes.RNA sequencing(RNA Seq)analysis showed differentially expressed genes(DEGs).Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were carried out to analyze enriched functions and signaling pathways.RT-qPCR,immunofluorescence,and western blotting were carried out to validate downstream gene expressions.The effects of tumorigenesis were evaluated in BALB/c nude mice by tumor xenografts.Our data revealed that the silencing of KONQ1OT1 in HCT116 and SW480 cells slowed cell growth and decreased the number of cells in the G2/M phase.RNA-Seq analysis showed the data of DEGs enriched in various GO and KEGG pathways such as DNA replication and cell cyde.RT qPCR,immunofluorescence,and western blotting confirmed downstream CCNE2 and PCNA gene expressions.HCT116 siKCN cells signifcantly suppressed tumorigenesis in BALB/c nude mice.Our study suggests that lncRNA KCNQ1OT1 may provide a promising therapeutic strategy for colon cancer.