Plasmacytoid dendritic cells mediate the tolerogenic effect of CD8+regulatory T cells in a rat tolerant liver transplantation model

BackgroundTolerance is more easily induced in liver transplant models than in other organs; CD8⁺CD45RC^lowregulatory T cells (Tregs) have been shown to induce tolerance in heart allografts. Whether CD8⁺CD45RC^lowTregs could induce tolerance in a liver transplant model and how dendritic cells (DCs) mediate the CD8⁺CD45RC^lowTregs effect remains to be investigated.MethodsA rat liver transplantation model was established and used to test tolerance and acute rejection compared to control groups. Liver function and histopathological changes of allograft were examined by enzyme-linked immunosorbent assay (ELISA) and haematoxylin and eosin (H&E) staining, respectively. The distribution and proportion of CD8⁺CD45RC^lowTregs and plasmacytoid dendritic cells (pDCs) in the allografts and spleen were determined using flow cytometry. Cytokine secretion levels were determined using ELISA and real-time quantitative PCR (qRT-PCR).ResultsThe rat liver transplantation model was well established, with a success rate of 93.3% (28/30). The mean survival time of the tolerant and acute-rejection rats were 156 and 14 days, respectively. The proportions of CD8⁺CD45RC^lowTegs were higher in the allografts of tolerant rats than in those of acute-rejection rats (33.1 ± 4.3 and 12.4 ± 4.6, respectively; P = 0.04). Significant accumulation of pDCs was observed in tolerant liver graft rats compared to that in acute-rejection rats (1.46 ± 0.23 and 0.80 ± 0.20, respectively; P = 0.02). Importantly, CD8⁺CD45RC^lowTregs were positively associated with the frequency of pDCs (P = 0.001, r² = 0.775). The protein and mRNA expression of IL-10 and TGF-β in the allograft group were increased, possibly being responsible for tolerance induction.ConclusionCD8⁺CD45RC^lowT cells interact with pDCs through the induction of IL-10 and TGF-β expression and are responsible for inducing immune tolerance in rat liver transplantation.     

心衰宁合剂对慢性心力衰竭大鼠心肌AMPK和PPARα的影响

目的探讨心衰宁合剂对慢性心力衰竭(chronic heart failure,CHF)大鼠心肌组织腺苷酸活化蛋白激酶(adenylate-activated protein kinase,AMPK)及过氧化物酶体增殖物激活受体α(peroxisome proliferator-activated receptorα,PPARα)表达的影响。方法将SD大鼠随机分为正常组,模型组,卡托普利组(10 mg·kg^-1),心衰宁合剂高、中、低剂量组(62.4、31.2、15.6 g·kg^-1)。正常组注射等量生理盐水,余组采用阿霉素(2.5 mg·kg^-1)腹腔注射制备慢性心力衰竭大鼠模型,每周1次。造模6周结束后,正常组及模型组给予生理盐水,余各组分别给予药物干预,干预4周。干预结束后,以HE染色观察心肌细胞形态学变化;Masson染色明确心肌纤维化程度;ELISA法检测大鼠血清B型钠尿肽(BNP)水平;比色法检测大鼠血清和心肌游离脂肪酸(FFA)水平;qRT-PCR、Western Blot法检测大鼠心肌AMPK、PPARα基因及蛋白表达水平。结果与正常组比较,模型组血清BNP水平明显升高(P<0.01),血清和心肌FFA水平明显升高(P<0.01),AMPK基因及蛋白表达明显升高(P<0.01),PPARα基因及蛋白表达明显降低(P<0.01);与模型组比较,心衰宁合剂各剂量组均降低血清BNP水平(P<0.01)及血清和心肌FFA水平(P<0.01,P<0.05),心衰宁合剂中、高剂量组可降低AMPK基因及蛋白表达水平(P<0.01,P<0.05),还可升高PPARα基因及蛋白表达水平(P<0.01)。结论心衰宁合剂可以减轻心力衰竭大鼠BNP含量,降低心衰大鼠心肌能量代谢底物FFA水平,其机制可能与调节PPARα、AMPK的蛋白与基因表达水平有关。     

单通道与单侧双通道脊柱内镜下腰椎间融合术治疗单节段腰椎退行性疾病的早期疗效及学习曲线

【摘要】 目的：比较单通道脊柱内镜下腰椎间融合术（endoscopic lumbar interbody fusion，Endo-LIF）与单侧双通道脊柱内镜下腰椎间融合术（unilateral biportal endoscopic lumbar interbody fusion，UBE-LIF）治疗单节段腰椎退行性疾病（lumbar degenerative diseases，LDD）的早期临床疗效，分析两者的学习曲线。方法：回顾性分析2019年10月~2022年3月在贵州医科大学附属医院骨科开展UBE-LIF术式与Endo-LIF术式治疗的54例单节段LDD患者，其中男26例，女28例；年龄55~71岁（65.0±4.5岁）；手术节段：L3/4 6例，L4/5 29例，L5/S1 19例。根据手术方式，分为A组（Endo-LIF治疗，n=24）和B组（UBE-LIF治疗，n=30）。根据患者接受手术时间的先后顺序，A组前12例为A1组，后12例为A2组；B组前15例为B1组，后15例为B2组。记录手术时间、术后住院时间及并发症，术前、术后3个月、6个月及1年时的腰腿痛视觉模拟评分（visual analogue scale，VAS）及Oswestry功能障碍指数（Oswestry disability index，ODI），末次随访时的改良Macnab标准评分及融合情况，计算优良率及融合率。通过曲线回归分析方法分析Endo-LIF与UBE-LIF手术时间随手术例数变化趋势。结果：所有患者手术均顺利完成，随访时间12~14个月（12.5±0.7个月）。与A2组相比，A1组手术时间较长（P<0.001），但术后住院时间两组间差异无统计学意义（P>0.05）。A组手术时间显著低于B1、B2组（P<0.001），且B1组明显大于B2组（P<0.001）。术后住院时间A组少于B1组（P<0.001），B2组与A组、B1组差异均无统计学意义（P>0.05）。术后A1组1例硬脊膜撕裂、B1组2例硬脊膜撕裂，均保守治疗后症状缓解；B1组1例发生类脊髓高压症，可能与手术时间较长、冲洗盐水刺激等有关，卧床休息后好转。A2与B2组未发生相关并发症。两组术后各时间点腰腿痛VAS评分及ODI较术前均显著改善（P<0.001），评分随时间推移较前下降（P<0.001）。末次随访时A组改良MacNab标准优良率为91.7%（22/24），B组为90.0%（27/30）；A组融合率87.5%，B组融合率90.0%。曲线回归分析示A、B组手术时间分别在12、15例手术后达到相对稳定。结论：Endo-LIF与UBE-LIF治疗LDD是安全有效的。随着手术例数增加两种术式的手术时间逐渐减少，Endo-LIF与UBE-LIF手术时间分别在12、15例手术后趋于稳定。     

3岁及以上人群新型冠状病毒灭活疫苗接种反应临床观察分析

目的 评价贵州省≥3岁人群接种新型冠状病毒（新冠病毒）灭活疫苗的安全性。方法 采用开放式设计，于2021年6月至2022年7月在贵州省沿河县招募符合条件的人群，按照（0，28）d免疫程序接种2剂次新冠病毒灭活疫苗，收集每剂次疫苗接种后30 min内和0~28 d的不良反应，分年龄、剂次、患病情况分析不良反应发生率。结果 总不良反应发生率为1.51%（294/19 458），所有不良反应均发生在接种后7 d内，严重程度为1级或2级，以接种部位疼痛最常见。3~、18~和≥60岁组不良反应发生率分别为1.01%（58/5 721）、2.44%（220/9 017）和0.34%（16/4 720），差异有统计学意义（P<0.001）。第1剂次接种后不良反应发生率（1.20%，233/19 458）高于第2剂次（0.37%，61/16 368），差异有统计学意义（P<0.001）。≥60岁组中健康人群和患基础疾病人群不良反应发生率分别为0.36%（9/2 520）和0.32%（7/2 200），差异无统计学意义（P=0.818）。结论 ≥3岁人群接种新冠病毒灭活疫苗安全性良好，在老年健康人群和患基础疾病人群中接种均具有良好安全性。     

Transcriptional changes in orthotopic liver transplantation and ischemia/reperfusion injury

BackgroundThere are few effective targeting strategies to reduce liver ischemia-reperfusion injury (IRI), which is one of the reasons for the poor prognosis of liver transplant recipients.MethodsA systematic approach combining gene expression with protein interaction (PPI) network was used to screen the characteristic genes and related biological functions of post-transplant. Differentially expressed genes (DEGs) between IRI+ and IRI- were identified. Logistic regression model and receiver operating characteristic (ROC) curve were used to identify potential target genes of IRI. The expression of key genes was verified by qRT-PCR and Western-blot experiments. Finally, the ssGSEA was used to identify the immune cell infiltration in patients with IRI.ResultsThe 283 common DEGs in GSE87487 and GSE151648 were mainly related to apoptosis and IL-17 signaling pathway. Through PPI network and logistic regression analysis, we identified that IL6, CCL2 and CXCL8 may be involved in the ischemia/reperfusion (IR) process. In addition, 32 genes were showed associated with IRI through inflammatory and metabolic pathways. Among the key genes identified, the differential expression of AGBL4, CILP2 and IL4I1 was verified by molecular experiments. Th17 cells of differentially infiltrated immune cells were positively correlated with CILP2 and IL4I1. The difference of Th17 cells between IRI+ and IRI- was verified by flow cytometry.ConclusionThe study showed that AGBL4, CILP2 and IL4I1 were associated with IRI. Th17 cells may be associated with the regulation of IRI by key genes. These genes and related pathways may be targets for improving IRI.     

Molecular mechanism of the ESRRG-PERM1-CKMT2 signal axis in ovariectomized female rats with OSAHS

This study aims to investigate the role of ESRRG-PERM1-CKMT2 signal axis, estrogen and genioglossal muscle contractile times in the development of Obstructive sleep apnea-hypopnea syndrome (OSAHS). This is a randomized controlled trial, and a total of 48 female rats are included and divided into following four groups: control group, sham operation group, model group and treatment group. A OSAHS rat model is established and then ESRRG, PERM1 and CKMT2 protein are detected by western blotting. Genioglossal muscle contractile times and estrogen level are also by detected. Our outcomes show that the level of ESRRG, PERM1, CKMT2 protein in control group, sham operation group, and treatment group are significantly up-regulated compared with that in model group. Genioglossal muscle contractile times and estrogen level in control group, sham operation group, and treatment group are also remarkably higher than those in model group. In addition, compared to the number of OSAHS in model group, the number of OSAHS in control group, sham operation group and treatment group is obviously decreased. We can conclude that up-regulated of ESRRG, PERM1, CKMT2 and estrogen can prevent or inhibit the development of OSAHS.     

徐学义教授治疗不寐临证经验拾萃

介绍徐学义教授治疗不寐的经验。其认为不寐的总纲在于阴阳失调,主要病机则为肝血不足、郁火扰神,主张在辨证施治的基础上,以交通阴阳、解郁清热、养血安神为基本诊疗原则,采用自拟方清解养血安神汤进行化裁结合特色药对治疗失眠症,效如桴鼓。     

Circ_0000274 contributes to renal cell carcinoma progression by regulating miR-338-3p/NUCB2 axis and JAK1/STAT3 pathway

BackgroundKidney transplant recipients (KTRs) are at increased risk of developing renal cell carcinoma (RCC). Accumulating evidence has demonstrated that circular RNAs (circRNAs) are essential players in tumor advancement. However, the functions of circ_0000274 in renal cell carcinoma (RCC) are barely explored.MethodsThe primary RCC cell lines 786-O and A498 were used in this study. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was employed for the RNA levels of circ_0000274, microRNA-338-3p (miR-338-3p) and nucleobindin 2 (NUCB2). RNase R assay was conducted to analyze the feature of circ_0000274.Cell Counting Kit-8 (CCK-8) assay, colony formation assay, transwell assay, tube formation assay and flow cytometry analysis were conducted for cell viability, colony formation, metastasis, angiogenesis and apoptosis, respectively. Western blot assay was utilized for protein levels. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were adopted to analyze the associations of circ_0000274 RNA, miR-338-3p RNA and NUCB2 protein. Murine xenograft model was established to explore the function of circ_0000274 RNA in vivo. Immunohistochemistry (IHC) assay was used to analyze NUCB2 protein level in xenograft tumors.ResultsCompared to normal tissues and cells, circ_0000274 RNA level was elevated in RCC tissues and cells. Knockdown of circ_0000274 RNA suppressed cell viability, colony formation, metastasis and tube formation and promoted apoptosis in RCC cells in vitro. Circ_0000274 RNA sponged miR-338-3p RNA to positively regulate NUCB2 protein in RCC cells. Inhibition of miR-338-3p reversed the impacts of circ_0000274 knockdown on RCC cell malignant behaviors. MiR-338-3p RNA overexpression repressed the malignant phenotypes of RCC cells, while NUCB2 protein elevation could abrogate the effect. Moreover, circ_0000274 RNA knockdown blocked tumorigenesis in vivo. Besides, circ_0000274 RNA knockdown inactivated the JAK1/STAT3 protein signaling pathway.ConclusionCirc_0000274 RNA functioned as an oncogene in RCC development by regulating miR-338-3p RNA/NUCB2 protein axis and activating the JAK1/STAT3 protein signaling pathway.     

浅析“依脉定腧”取穴法的临床应用＊

腧穴是人体脏腑经络之气输注出入的特殊部位，既是疾病的反应点，又是针灸防治疾病的刺激点，其主要存在“静息”和“敏化”两种状态。在临床上，本课题组依据腧穴在疾病状态下的力敏效应来选取针刺穴位，经过多年临床观察发现，对患者进行经络诊察的过程中，应用“循、按、扪、切”等操作手法刺激到人体某一个部位时，会即刻引起患者寸口气口九道脉中的病变经脉脉象发生改变，且该脉象的变化可随刺激的停止而消失，继续而重现。本课题组根据《易经》中“寂然不动，感而遂通”的理论观点，再加上理论上医源于易，医易本属一家，所以就二者间存在的相似之处，临床可类比应用。本研究结合自身对该理论观点的理解，通过临床经络诊察刺激患者在疾病状态下的敏化腧穴，引起寸口气口九道脉中病变经脉脉象发生改变的临床实践方式，选取针刺穴位，收效颇著。因此，本研究提出了“依脉定腧”的新型取穴方法，指明了引起脉象变化的特殊部位即为患者在生病状态下的敏化腧穴，是针刺治疗起效的关键穴位。     

长春新碱诱导的周围神经病变的儿童队列研究

目的 总结急性淋巴细胞白血病（acute lymphoblastic leukemia,ALL）患儿发生长春新碱诱导的周围神经病变（vincristine-induced peripheral neuropathy,VIPN）的特征，探讨VIPN发生的影响因素。方法 选择2018年1月—2022年2月于贵州医科大学附属医院使用CCCG-ALL2015或CCCG-ALL2020方案治疗的1～18岁ALL患儿为研究对象，根据治疗方案中年龄对危险度分组的影响，将患儿分为1～10岁组（91例）和>10岁组（29例），并根据国家癌症研究所不良事件通用术语标准第5版，对发生的VIPN进行分级评估，比较不同组间在VIPN发生率、严重程度、类型上的差异。结果 共纳入120例患儿，56例发生VIPN，发生率为46.7%。>10岁组患儿VIPN发生率高于1～10岁组（69%vs 40%,P<0.05）。在发生VIPN患儿中，3级及以上VIPN 12例（21%）,2级VIPN 44例（79%）。自主神经症状77例次（59.7%）、周围神经损伤42例次（32.5%）、颅神经损伤10例次（7....