C-X-C chemokine receptor type 5+CD8+T cells as immune regulators in hepatitis Be antigen-positive chronic hepatitis B under interferonalpha treatment

BACKGROUND C-X-C chemokine receptor type 5(CXCR5)+CD8+T cells represent a unique immune subset with dual roles,functioning as cytotoxic cells in persistent viral infections while promoting B cell responses.Despite their importance,the specific role of CXCR5+CD8+T cells in chronic hepatitis B(CHB),particularly during interferon-alpha(IFN-α)treatment,is not fully understood.This study aims to elucidate the relationship between CXCR5+CD8+T cells and sustained serologic response(SR)in patients undergoing 48 weeks of pegylated IFN-α(peg-IFN-α)treatment for CHB.AIM To elucidate the relationship between CXCR5+CD8+T cells and sustained SR in patients undergoing 48 weeks of peg-IFN-αtreatment for CHB.METHODS This study enrolled 60 patients with hepatitis Be antigen(HBeAg)-positive CHB undergoing 48 weeks of peg-IFN-αtreatment.Participants were assessed for eligibility based on criteria such as persistent HBsAg-positive status for at least six months,HBeAb-negative,hepatitis B virus DNA levels exceeding 2×104 copies/mL,and alanine aminotransferase(ALT)levels between 2 and 10 times the upper limit of normal.Blood samples were collected at baseline and at weeks 12,24,48,and a 24-week treatment-free follow-up(week 72)to measure serum interleukin(IL)-21 concentration via ELISA and to analyze CXCR5 and programmed death-ligand 1(PD-L1)expression on CD8+T cells by flow cytometry,CXCR5 is a chemokine receptor that directs immune cells to specific tissues,while PD-L1 is a protein that regulates immune responses by inhibiting T cell activity.RESULTS Patients with CHB exhibited significantly lower levels of circulating CXCR5+CD8+T cells compared to healthy controls(P<0.01).Notably,CXCR5+CD8+T cells were prominently expressed in patients who achieved sustained SR compared to non-SR(NSR).A significant correlation was observed between CXCR5 and PD-L1 expression(r=-0.189,P=0.002).However,there was no significant correlation between serum IL-21 levels and CXCR5+CD8+lymphocytes(r=-0.03,P=0.625)or serum ALT levels(r=0.026,P=0.678).CONCLUSION The enhanced expression of CXCR5+CD8+T cells in patients achieving HBeAg seroconversion during IFN-αtreatment suggests that these cells play a crucial role in antiviral immune responses against hepatitis B.This study highlights the potential of CXCR5+CD8+T cells as immune regulators in CHB,which may inform future therapeutic strategies to optimize antiviral treatments.

Augmentative effect of tetrandrine on pentobarbital hypnosis mediated by 5-HT_(1A) and 5-HT_(2A/2C) receptors in mice

It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.

慢性阻塞性肺疾病急性加重期患者血清CCR5、PAI-1水平与肺功能及病情严重程度的关系

目的 探究慢性阻塞性肺疾病急性加重期(AECOPD)患者血清细胞趋化因子受体5(CCR5)、纤溶酶原激活物抑制剂-1(PAI-1)水平变化与肺功能及病情严重程度之间的关系。方法 选取该院2020年1月至2022年1月急诊收治的114例AECOPD患者为AECOPD组,并选取同期稳定期慢性阻塞性肺疾病(COPD)患者114例为对照组。采用酶联免疫吸附试验(ELISA)测定CCR5、PAI-1水平;比较AECOPD组与对照组及不同病情程度AECOPD患者血清CCR5、PAI-1水平及肺功能指标;Pearson相关性分析AECOPD患者血清CCR5、PAI-1水平与肺功能指标之间的关系;Spearman相关性分析AECOPD患者血清CCR5、PAI-1水平与病情严重程度之间的关系。结果 与对照组相比,AECOPD组患者血清CCR5、PAI-1水平均显著升高,差异均有统计学意义(P<0.05),肺功能指标:第1秒用力呼气量(FEV_(1))、FEV_(1)与用力肺活量比值(FEV_(1)/FVC)、FEV_(1)占预计值百分比(FEV_(1)%pred)、最大呼气中期流量(MMEF)占预计值百分比(MMEF%pred)均显著降低,差异有统计学意义(P<0.05),且随着病情程度的增加,血清CCR5、PAI-1水平逐渐增加(P<0.05),FEV_(1)、FEV_(1)/FVC、FEV_(1)%pred、MMEF%pred水平逐渐降低(P<0.05);Pearson相关性分析结果显示,血清CCR5、PAI-1水平与FEV_(1)、FEV_(1)/FVC、FEV_(1)%pred、MMEF%pred均呈显著负相关(P<0.05);Spearman相关性分析结果显示,AECOPD患者血清CCR5、PAI-1水平与病情程度呈显著正相关(r_(1)=0.432,r_(2)=0.451,P<0.05)。结论 AECOPD患者血清CCR5、PAI-1水平显著升高,且血清CCR5、PAI-1水平与AECOPD患者肺功能及病情程度密切相关。

S1PR5对脂多糖诱导小鼠认知行为和炎症反应的影响及其抗炎机制

目的:探讨鞘氨醇1-磷酸受体5(S1PR5)对脂多糖(LPS)诱导的神经炎症、认知行为的影响,及对BV2细胞的抗炎作用及相关机制。方法:(1)使用C57BL/6野生型(WT)小鼠和同背景S1PR5基因敲除(knockout,KO)小鼠,按照随机数字法分为WT对照组、WT-LPS组、S1PR5 KO对照组和S1PR5 KO-LPS组。WT-LPS组和S1PR5 KO-LPS组小鼠分别经单次腹腔注射5 mg/kg LPS构建神经炎症模型。WT对照组和S1PR5 KO对照组注射等量生理盐水。造模7 d后进行Morris水迷宫实验,之后收集各组小鼠脑组织,尼式染色观察海马组织变化;RT-qPCR检测海马组织中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-6的mRNA表达水平。Western blot和组织免疫荧光检测海马组织中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)的表达。(2)以LPS刺激BV2细胞诱导炎症反应并用A971432激动S1PR5或慢病毒过表达S1PR5进行干预,通过RT-qPCR法分别检测激动或过表达S1PR5对S1PR5、IL-1β、IL-6、TNF-α及CD206的影响;细胞免疫荧光法检测CD206的表达;Western blot法检测CD206、诱导型一氧化氮合酶(iNOS)、环加氧酶2(COX2)、NLRP3、p-NF-κB、cleaved caspase-1和IκBα的蛋白水平。结果:(1)体内实验发现,S1PR5 KO可显著增加LPS诱导的小鼠记忆缺损、海马区IL-1β和IL-6的mRNA表达及NLRP3蛋白表达(P<0.05或P<0.01)。(2)BV2细胞中S1PR5的表达不具有A971432浓度依赖性及时间依赖性。(3)激动S1PR5或过表达S1PR5可在mRNA水平上显著降低LPS诱导BV2细胞表达IL-1β、IL-6和TNF-α,增加CD206的表达(P<0.05或P<0.01);可在蛋白水平上显著增加M2型巨噬细胞标志蛋白CD206,降低M1型巨噬细胞标志物iNOS和COX2的表达(P<0.05或P<0.01);可显著降低NLRP3、p-NF-κB和cleaved caspase-1的表达,增加IκBα的表达(P<0.05或P<0.01)。结论:(1)S1PR5缺失通过促进神经炎症反应加重了LPS诱导的小鼠认知障碍。(2)激动或过表达S1PR5促进BV2细胞向M2表型极化,抑制NLRP3炎症小体组成成分的表达及NF-κB信号通路,抑制LPS诱导的炎症反应。

TypeⅠinositol 1, 4, 5-triphosphate receptors increase in kidney of mice with fulminant hepatic failure

AIM： To delineate the mechanisms of renal vasoconstriction in hepatorenal syndrome （HRS）, we investigated the expression of type I inositol 1, 4, 5-triphosphate receptors （IP3R I） of kidney in mice with fulminant hepatic failure （FHF）. METHODS： FHF was induced by lipopolysaccharide （LPS） in D-galactosamine （GAIN） sensitized BALB/c mice. There were 20 mice in normal saline （NS）-treated group, 20 mice in LPS-treated group, 20 mice in GaIN- treated group, and 60 mice in GalN/LPS-treated group （FHF group）. Liver and kidney tissues were obtained at 2, 6, and 9 h after administration. The liver and kidney specimens were stained with hematoxylin-eosin for studying morphological changes under light microscope. The expression of IP3R I in kidney tissue was tested by immunohistochemistry, Western blot and reverse transcription （RT）-PCR. RESULTS： Kidney tissues were morphologically normal at all time points in all groups. IP3R I proteins were found localized in the plasma region of glomerular mesangial cells （GMC） and vascular smooth muscle cells （VSMC） in kidney by immunohistochemical staining. In kidney of mice with FHF at 6 h and 9 h IP3R I staining was upregulated. Results from Western blot demonstrated consistent and significant increment of IP3R I expression in mice with FHF at 6 h and 9 h （t = 3.16, P 〈 0.05; t = 5.43, P 〈 0.01）. Furthermore, we evaluated IP3R I mRNA expression by RT-PCR and observed marked upregulation of IP3R I mRNA in FHF samples at 2 h, 6 h and 9 h compared to controls （t = 2.97, P 〈 0.05; t = 4.42, P 〈 0.01; t = 3.81, P 〈 0.01）. CONCLUSION： The expression of IP3R I protein increased in GMC and renal VSMC of mice with FHF, possibly caused by up-regulation of IP3R I mRNA.

LncRNA AFAP1-AS1 exhibits oncogenic characteristics and promotes gemcitabine-resistance of cervical cancer cells through miR-7-5p/EGFR axis

Background:Drug resistance is the main factor contributing to cancer recurrence and poor prognosis.Exploration of drug resistance-related mechanisms and effective therapeutic targets are the aim of molecular targeted therapy.In our study,the role of long non-coding RNA(lncRNA)AFAP1-AS1 in gemcitabine resistance and related mechanisms were explored in cervical cancer cells.Methods:Gemcitabine-resistant cervical cancer cell lines HT-3-Gem and SW756-Gem were constructed using the gemcitabine concentration gradient method.The overall survival rates and recurrence-free survival rates were evaluated by Kaplan-Meier analysis.The interaction was verified through a Dual-luciferase reporter gene assay and a Biotinylated RNA pull-down assay.Cell proliferation ability was assessed through methyl-thiazolyl-tetrazolium(MTT),soft agar,and colony formation experiments.Cell cycle and apoptosis were detected byflow cytometry.Results:Up-regulation of AFAP1-AS1 in cervical cancer predicted a poor prognosis.Besides,patients in the gemcitabine-resistance group had higher levels of AFAP1-AS1 than the gemcitabine-sensitive group.AFAP1-AS1 promoted tumor growth and induced gemcitabine tolerance of cervical cancer cells.In addition,AFAP1-AS1 mediated epidermal growth factor receptor(EGFR)expression by serving as a molecular sponge for microRNA-7a-5p(miR-7-5p).This present study also proved that the knockdown of EGFR or overexpression of miR-7a-5p abolished the accelerative role of AFAP1-AS1 overexpression in cancer progression and gemcitabine tolerance.Conclusions:In general,the AFAP1-AS1/miR-7-5p/EGFR axis was tightly related to the progression and gemcitabine tolerance of cervical cancer,providing potential targets for the management of cervical cancer.

Therapeutic Trial of an Endothelin Receptor Agonist for the Highly Pathogenic Avian Influenza A/H5N1 Virus Infection in Chicks

The rapid spread of the highly pathogenic A/H5N1 avian influenza virus among domestic birds and its transmission to humans has induced world-wide fears of a new influenza pandemic. A/H5N1 has infected over 300 people since 1997, and has shown a mortality rate of over 50%. The high mortality in human cases is thought to be enhanced by the excessive secretion of various endogenous factors, including cytokines and interleukins, stimulated by viral infections. Chickens infected with A/H5N1 viruses experience sudden death without showing severe clinical symptoms or inflammation. However, severe hemorrhage and congestion are seen in various tissues in sporadic chicken cases of A/H5N1-infections, especially in the pulmonary tissues, thus indicating that there is ischemia due to vascular abnormalities. Our previous studies have focused on the expression pattern of endothelin-1, which modulates the vascular tone via endothelin receptors. An Indonesian sporadic strain of A/H5N1 virus was intranasally administered to 10-day-old chicks, and the expression of endothelin was examined in the infected birds. All birds died within five days of inoculation, and had moderate inflammation accompanied by severe hemorrhage and congestion in the lungs. Immunohistochemical studies showed enhanced expression of endothelin-1 in the infected lungs. In addition, the real-time PCR analyses revealed that endothelin-1 and endothelin receptor A mRNA were significantly elevated in the birds with A/H5N1 infections. Subsequently, H5N1-infected birds were inoculated with bosentan hydrate, a competitive antagonist of endothelin receptors. Interestingly, the mortality rate of the infected birds was dramatically decreased in a dose-dependent manner by the administration of bosentan hydrate. The pathological lesions, including congestion and hemorrhage in the pulmonary tissues, were clearly inhibited. These findings are promising, and suggest that endothelin receptor antagonists are a potential treatment for the highly pathogenic avian flu.

Mutations in Hemagglutinin of H5N1 Influenza That Switch Receptor Specificity from Avian to Human Types

Researchers have been searching for molecular features that could make avian H5N1 influenza transmissible among people since the first report of human infections with this virus in 1997. A recent study surprisingly demonstrated that only five mutations, fewer than previously estimated, are needed to make avian H5N1 influenza transmissible between ferrets through the air, raising fears that a human pandemic is possible if this virus escapes from the lab. Of the five mutations found, four of them are located in the HA gene that is responsible for the viral entry into the host cells. A crucial step for avian influenza to go across the species boundary to infect humans is the switch of its receptor binding specificity from avian to human types. The first task of this study was to quantify the individual as well as the collective effect of the known HA mutations from the previous research on receptor binding selection. Our second task was to identify new combinations of HA mutations that could change the receptor binding preference of H5N1 from avian to human types. Our findings thus deepened our understanding of the previous research and also extended its results by discovering new combinations of mutations that could enhance the binding of avian H5N1 to human type receptors while reduce that to avian types.

复发性口腔溃疡患儿血清IRF5、sTREM-1变化及对复发的预测价值

目的探讨复发性口腔溃疡患儿血清干扰素调节因子5(IRF5)、可溶性髓样细胞触发受体1(sTREM-1)水平变化及其对复发的预测价值。方法选取石家庄市妇幼保健院于2021年7月至2022年12月收治的146例复发性口腔溃疡患儿为观察组,并根据3个月内是否复发分为复发组(n=45)和未复发组(n=101)。另选取同期138例健康志愿者作为对照组。采用实时荧光定量PCR(qPCR)检测血清IRF5 mRNA表达水平,酶联免疫吸附试验(ELISA)检测血清sTREM-1、白细胞介素(IL)-6和IL-10表达水平,Pearson分析法分析复发性口腔溃疡患儿血清IRF5和sTREM-1水平与IL-6和IL-10水平的相关性,受试者工作特征(ROC)曲线分析血清IRF5和sTREM-1水平对复发性口腔溃疡患儿复发的预测价值。结果与对照组比较,观察组血清IRF5水平明显降低(P<0.05),sTREM-1、IL-6和IL-10水平明显升高(P<0.05);Pearson分析结果显示,复发性口腔溃疡患儿血清IRF5与IL-6、IL-10水平呈负相关(r=-0.531、-0.462,P<0.05),血清sTREM-1水平与IL-6、IL-10水平呈正相关(r=0.435、0.480,P<0.05);与未复发组比较,复发组患儿血清IRF5水平明显降低(P<0.05),sTREM-1水平明显升高(P<0.05)。ROC曲线分析结果显示,血清IRF5水平单独预测复发性口腔溃疡患儿复发的曲线下面积(AUC)为0.823,最佳cut-off值为0.85,灵敏度和特异度分别为75.56%、84.16%;血清sTREM-1水平单独预测复发性口腔溃疡患儿复发的AUC为0.833,灵敏度和特异度分别为68.89%、88.12%,最佳cut-off值为84.08 pg/mL;二者联合预测复发性口腔溃疡患儿复发的灵敏度和特异度分别为88.89%、82.18%,AUC为0.915,显著高于IRF5单独预测的AUC(Z=2.455,P=0.014)和sTREM-1单独预测的AUC(Z=2.790,P=0.005)。结论复发性口腔溃疡患儿血清IRF5水平较低、sTREM-1水平较高,二者联合对复发性口腔溃疡患儿再复发具有较高的预测价值。

GPRC5A调控的ABCB1表达对肺腺癌增殖的影响

目的ATP结合盒B亚家族成员1(ATP binding cassette subfamily B member 1,ABCB1)的异常表达在多种癌症的发生发展中发挥关键作用。然而,G蛋白偶联受体C家族5组A型(G protein coupled receptor family C group5 type A,GPRC5A)调控的ABCB1表达对肺腺癌增殖的影响仍不清楚。本研究探讨了GPRC5A调控的ABCB1表达对肺腺癌增殖的影响。方法我们采用RT-PCR、Western-blot或免疫组化实验,分析ABCB1在肺腺癌细胞系、人肺腺癌组织以及GPRC5A基因敲除小鼠和野生型小鼠的气管上皮细胞和肺组织中的表达。采用细胞计数试剂盒-8(CCK-8)分析GPRC5A基因敲除小鼠气管上皮细胞对化疗药物的敏感性。采用皮下肿瘤形成实验探讨下调ABCB1表达是否可抑制体内肺腺癌增殖。采用免疫荧光和免疫沉淀实验研究GPRC5A和ABCB1之间潜在的调控关系。结果ABCB1在肺腺癌细胞系和人类肺腺癌组织中表达上调。GPRC5A基因敲除小鼠的气管上皮细胞及肺组织的ABCB1表达高于野生型小鼠。与GPRC5A野生型小鼠的气管上皮细胞相比,GPRC5A基因敲除小鼠的气管上皮细胞对塔立奇达和多柔比星更敏感。注射移植细胞28天后,接受ABCB1基因敲除细胞移植的GPRC5A-/-C57BL/6小鼠的肺肿瘤的体积和重量均明显低于野生型细胞移植小鼠(P=0.0043,P=0.0060)。此外,免疫荧光和免疫沉淀实验表明,GPRC5A通过直接结合方式调控ABCB1的表达。结论GPRC5A通过抑制ABCB1表达降低肺腺癌增殖。GPRC5A调节ABCB1表达的途径有待研究。

Netrin-1 signaling pathway mechanisms in neurodegenerative diseases

Netrin-1 and its receptors play crucial roles in inducing axonal growth and neuronal migration during neuronal development.Their profound impacts then extend into adulthood to encompass the maintenance of neuronal survival and synaptic function.Increasing amounts of evidence highlight several key points:(1)Diminished Netrin-1 levels exacerbate pathological progression in animal models of Alzheimer’s disease and Parkinson’s disease,and potentially,similar alterations occur in humans.(2)Genetic mutations of Netrin-1 receptors increase an individuals’susceptibility to neurodegenerative disorders.(3)Therapeutic approaches targeting Netrin-1 and its receptors offer the benefits of enhancing memory and motor function.(4)Netrin-1 and its receptors show genetic and epigenetic alterations in a variety of cancers.These findings provide compelling evidence that Netrin-1 and its receptors are crucial targets in neurodegenerative diseases.Through a comprehensive review of Netrin-1 signaling pathways,our objective is to uncover potential therapeutic avenues for neurodegenerative disorders.

Prolonged intermittent theta burst stimulation restores the balance between A_(2A)R-and A_(1)R-mediated adenosine signaling in the 6-hydroxidopamine model of Parkinson's disease

An imbalance in adenosine-mediated signaling,particularly the increased A_(2A)R-mediated signaling,plays a role in the pathogenesis of Parkinson's disease.Existing therapeutic approaches fail to alter disease progression,demonstrating the need for novel approaches in PD.Repetitive transcranial magnetic stimulation is a non-invasive approach that has been shown to improve motor and non-motor symptoms of Parkinson's disease.However,the underlying mechanisms of the beneficial effects of repetitive transcranial magnetic stimulation remain unknown.The purpose of this study is to investigate the extent to which the beneficial effects of prolonged intermittent theta burst stimulation in the 6-hydroxydopamine model of experimental parkinsonism are based on modulation of adenosine-mediated signaling.Animals with unilateral 6-hydroxydopamine lesions underwent intermittent theta burst stimulation for 3 weeks and were tested for motor skills using the Rotarod test.Immunoblot,quantitative reverse transcription polymerase chain reaction,immunohistochemistry,and biochemical analysis of components of adenosine-mediated signaling were performed on the synaptosomal fraction of the lesioned caudate putamen.Prolonged intermittent theta burst stimulation improved motor symptoms in 6-hydroxydopamine-lesioned animals.A 6-hydroxydopamine lesion resulted in progressive loss of dopaminergic neurons in the caudate putamen.Treatment with intermittent theta burst stimulation began 7 days after the lesion,coinciding with the onset of motor symptoms.After treatment with prolonged intermittent theta burst stimulation,complete motor recovery was observed.This improvement was accompanied by downregulation of the e N/CD73-A_(2A)R pathway and a return to physiological levels of A_(1)R-adenosine deaminase 1 after 3 weeks of intermittent theta burst stimulation.Our results demonstrated that 6-hydroxydopamine-induced degeneration reduced the expression of A_(1)R and elevated the expression of A_(2A)R.Intermittent theta burst stimulation reversed these effects by restoring the abundances of A_(1)R and A_(2A)R to control levels.The shift in ARs expression likely restored the balance between dopamine-adenosine signaling,ultimately leading to the recovery of motor control.

Rivastigmine Restores 5-HT_1AReceptor Levels in the Hippocampus of Olfactory Bulbectomized Mice

Rivastigmine, a dual acetylcholinesterase and butyrylcholinesterase inhibitor, is used for symptomatic treatment of patients with mild to moderately severe dementia in Alzheimer’s disease (AD) patients. In the present study, we found that 5-HT1A receptor (5-HT1AR) is downregulated, whereas 5-HT2A receptor (5-HT2AR) is upregulated in the hippocampal dentate gyrus (DG) and CA1 region by olfactory bulbectomy (OBX) in mice. Furthermore, chronic treatment with rivastigmine (1.0 mg/kg) for 2 weeks starting 2 weeks after OBX operation restored the decreased 5-HT1AR and the increased 5-HT2AR levels. To determine whether cholinergic receptor stimulation by rivastigmine is involved in the rivastigmine-induced regulation of 5-HTR levels, we treated the mice with mecamylamine (2.5 mg/kg), or atropine (5.0 mg/kg) with rivastigmine (1.0 mg/kg) once a day for 2 weeks. Notably, the rivastigmine-induced 5-HT1AR upregulation was eliminated by mecamylamine but not by atropine treatments. On the other hand, the restored 5-HT2AR level by rivastigmine was not affected by either mecamylamine or atropine. Treatment with 8-OH-DPAT, a selective 5-HT1AR agonist improved the decreased 5-HT1AR and the increased 5-HT2AR levels in OBX mice. On the other hand, treatment with TCB-2, a potent 5-HT2AR agonist had no effects on the 5-HT1AR and 5-HT2AR dysregulation in OBX mice. Taken together, nicotinic acetylcholine receptor (nAChR) stimulation mediates rivastigmine-induced upregulation of 5-HT1AR. Therefore, we speculate that the increased ACh levels by rivastigmine can stimulate nAChR located on serotonergic nerve terminals and stimulate 5-HT1AR by the enhanced 5-HT release in the hippocampus. The 5-HT1AR stimulation likely mediates the improvement of 5-HT1AR levels as auto-receptor in OBX hippocampus.

Role of doublecortin-like kinase 1 and leucine-rich repeat-containing G-protein-coupled receptor 5 in patients with stage Ⅱ/Ⅲ colorectal cancer:Cancer progression and prognosis

BACKGROUND Cancer stem cells(CSCs)are a subpopulation of cancer cells with the potential of self-renewal and differentiation.CSCs play critical roles in tumorigenesis,recurrence,metastasis,radiation tolerance and chemoresistance.AIM To assess the expression patterns and clinical potential of doublecortin-like kinase 1(DCLK1)and leucine-rich repeat-containing G-protein-coupled receptor 5(Lgr5),as prognostic CSC markers of colorectal cancer(CRC).METHODS The expression of DCLK1 and Lgr5 in CRC tissue sections from 92 patients was determined by immunohistochemistry.Each case was evaluated using a combined scoring method based on signal intensity staining(scored 0-3)and the proportion of positively stained cancer cells(scored 0-3).The final staining score was calculated as the intensity score multiplied by the proportion score.Low expression of DCLK1 and Lgr5 was defined as a score of 0-3;high expression of DCLK1 and Lgr5 was defined as a score of≥4.Specimens were categorized as either high or low expression,and the correlation between the expression of DCLK1 or Lgr5 and clinicopathological factors was investigated.RESULTS DCLK1 and Lgr5 expression levels were significantly positively correlated.CRC patients with high DCLK1,Lgr5 and DCLK1/Lgr5 expressions had poorer progression-free survival and overall survival.Moreover,high expression of DCLK1 was an independent prognostic factor for recurrence and overall survival in patients with CRC by multivariate analysis(P=0.026 and P=0.049,respectively).CONCLUSION DCLK1 may be a potential CSC marker for the recurrence and survival of CRC patients.

Effects of Activation and Blockade of Serotonin 5-HT1A Receptors on the Immune Response in Rats Selected for Different Levels of Aggressiveness

The present study examines the effects of serotonin (5-HT) 1A receptor ligands on humoral im-mune response in two rat lines selected for over 75 generations for the enhancement or elimination of aggression. Activation of presynaptic 5-HT1A receptors with a low dose of the selective 5-HT1A receptor agonist 8-OH-DPAT (0.1 mg/kg) or the blockade of postsynaptic 5-HT1A receptors with the antagonist WAY-100635 (1.0 mg/kg) did not affect the numbers of IgM-antibody forming cells (IgM-AFC) in the spleen of highly aggressive rats, which were characterized by higher immune responsiveness compared to nonaggressive line. On the other hand, the same doses of 8-OH-DPAT and WAY-100635, as well as a higher dose of 8-OH-DPAT (1.0 mg/kg), which is known to activate postsynaptic 5-HT1A receptors, produce immunostimulation in nonaggressive rats. However, only the highest dose of 8-OH-DPAT (5.0 mg/kg) was able to cause immunosuppression in nonaggressive rats that was mainly dependent on stimulation of postsynaptic 5-HT1A receptors. In contrast to nonaggressive rats, the dose of 1.0 mg/kg 8-OH-DPAT was sufficient to produce a decrease in the numbers of IgM-AFC in highly aggressive rats. Thus, pharmacological activation of pre- and postsynaptic 5-HT1A receptors, as well as the blockade of postsynaptic 5-HT1A receptors, produced different effects on the immune response in two lines of rats selected for high level of aggression or its absence. These data may have implications for more efficient treatments of a number of mental disorders associated with abnormal aggression.

去卵巢大鼠体温变化与下丘脑视前区5-羟色胺_(1A)受体表达

目的观察大鼠去卵巢后体温变化与下丘脑视前区5-羟色胺1A(5-HT1A)受体亚型表达之间的关系。方法用数字温度计TM902C检测大鼠的肛温,酶联免疫吸附试验测定血清激素水平,根据5-HT1A受体互补DNA序列合成相应的特异性引物,用聚合酶链反应(PCR)法观察大鼠去卵巢后下丘脑视前区5-HT1A受体表达。结果 (1)大鼠去卵巢(OVX) 组自第8周末肛温明显高于假去势(sham)组(P<0．05)。(2)OVX组大鼠血清17β-雌二醇(17β-E2)较sham组下降,黄体生成素(LH)水平升高,差异显著(P<0．05)。(3)OVX组下丘脑视前区5-HT1A受体表达增强,与sham组比较有显著性差异(P <0．05)。结论大鼠卵巢摘除后体温升高,下丘脑视前区5-HT1A受体表达增强,提示5-HT在低雌激素状态下的体温调节中起一定作用。

直肠内脱垂患者直肠顺应性变化与肠黏膜中TRPV1、5-HT表达的关系

目的探讨直肠内脱垂(internal rectal prolapse,IRP)患者直肠顺应性改变与TRPV1、5-HT在直肠黏膜层表达的关系。方法选择临床诊断IRP的患者20例及对照组20例先进行肛管直肠测压,然后行结肠镜检查,在齿状线上方4 cm处下段直肠取黏膜活检,行TRPV1和5-HT免疫组化染色,采用图像分析法分析免疫反应阳性情况。结果与对照组比较,IRP组初始感觉容积明显增高(Z=-3.710,P<0.01),最大感觉耐受容积明显降低(Z=-5.181,P<0.01),直肠顺应性明显降低;对照组黏膜层仅有少量TRPV1表达,IRP组直肠黏膜腺体可见大量TRPV1阳性染色,与对照组相比,IRP组直肠黏膜层TRPV1的表达显著增强(Z=-5.085,P<0.01);对照组黏膜层5-HT免疫阳性染色很少,而IRP组黏膜层腺体细胞内5-HT表达量显著升高(Z=-4.734,P<0.01)。结论 IRP患者直肠顺应性明显低于对照组,与直肠黏膜层TRPV1和5-HT免疫反应阳性显著增强导致直肠高敏感有关,可能是IRP患者肛门坠胀、便意频繁等排便功能障碍的机制之一。

中国傣族、景颇族人群中与艾滋病相关的CCR5△32、CCR2b-64I、SDF1-3′A等位基因多态性分布

为了调查HIV 1感染相关的等位基因CCR5△32、CCR2b 64I、SDF1 3′A在我国云南省德宏州傣族景颇族人群中的频率和多态性分布,此课题以101例傣族和113例景颇族人群为研究对象,应用PCR、PCR RFLP(聚合酶链反应 限制性片段长度多态性)分析方法进行检测,计算突变基因频率;并对其群体分布、性别分布进行统计学分析。结果表明,中国傣族景颇族人群中未发现CCR5△32等位基因突变;傣族CCR2b 64I、SDF1 3′A基因突变频率分别为0.2130和0.2030,景颇族CCR2b 64I和SDF1 3′A基因突变频率分别为0.1637和0.1770;与中国汉族人群相比较,傣族和景颇族中SDF1 3′A突变频率较低(P值分别为0.0322和0.0021);两个民族的CCR2b 64I和SDF1 3′A等位基因群体分布符合Hardy Weinberg平衡,在性别之间分布无显著差异。中国傣族景颇族人群的CCR2b 64I等位基因的突变频率与汉族人相似,SDF1 3′A等位基因的突变频率比汉族人低,此两种突变基因在艾滋病发病过程中的影响值得进一步研究。由于未发现CCR5△32基因突变,中国傣族景颇族人群对HIV 1感染可能有较大的遗传易感性。

5-羟色胺1A受体基因多态性与精神分裂症关联性的Meta分析

目的:用Meta分析的方法综合评价人群中5-羟色胺1A(5-HT1A)受体基因多态性与精神分裂症的关联,为精神分裂症的遗传背景提供循证医学证据。方法:计算机检索PubMed、Cochorane、CNKI和万方数据库,搜集有关5-HT1A受体基因多态性与精神分裂症关联性的全文文献。以病例组和对照组5-HT1A受体基因分布的OR及其95%CI为效应指标,在全面文献回顾的基础上对文献进行筛选、评价和数据提取。应用RevMan5.0软件对各研究原始数据进行统计分析,包括异质性检验、合并效应量以及评估发表偏倚。结果:共有5篇文献被纳入分析,其中2篇研究rs6294位点与精神分裂症的关联性,3篇研究rs6295位点与精神分裂症的关联性。精神分裂症患者rs6294位点的基因型Meta分析结果,合并OR=0.35,95%CI=0.12～1.08,P=0.07;rs6294位点等位基因Meta分析结果,合并OR=0.34,95%CI=0.11～1.03,P=0.06;精神分裂症患者rs6295位点的基因型Meta分析结果,合并OR=1.78,95%CI=0.69～4.59,P=0.23;rs6295位点的等位基因Meta分析结果,合并OR=1.80,95%CI=1.03～3.16,P=0.04。结论:人群中5-HT1A受体基因多态性与精神分裂症有关联。

代谢型谷氨酸受体1和5激动剂改善阿霉素诱导的小鼠肾损伤

目的研究选择性代谢型谷氨酸受体1和5(mGluR1/5)激动剂3,5-二羟基苯甘氨酸(DHPG)对阿霉素(ADR)肾病小鼠模型的作用。方法 18只6周龄BALB/c小鼠随机分为对照组、ADR组(经尾静脉注射阿霉素建立阿霉素肾病小鼠模型)和ADR+DHPG组(建模前预先注射DHPG),每组6只。应用酶联免疫吸附试验(ELISA)方法检测小鼠24 h尿白蛋白水平,透射电子显微镜观察足细胞足突形态和宽度;免疫荧光染色激光共聚焦显微镜观察足细胞标志蛋白nephrin和损伤标志蛋白desmin的表达,免疫组织化学法检测肾小球WT-1表达;TUNEL染色检测足细胞凋亡情况。结果在对照组、ADR组和ADR+DHPG组24 h尿白蛋白分别为(1.67±0.22)、(22.55±2.34)和(8.23±2.74)mg,组间比较差异有统计学意义(P<0.01)。与ADR组比较,ADR+DHPG组小鼠足细胞的足突宽度缩短,desmin表达减少,而nephrin表达增加。肾小球中WT-1阳性细胞数与凋亡细胞数呈显著负相关(R2=0.8 482,P<0.05);ADR+DHPG组足细胞凋亡数显著少于ADR组(P<0.01)。结论选择性mGluR1/5激动剂DHPG可抑制ADR诱导的足细胞足突融合、nephrin表达减少和细胞凋亡,减少阿霉素肾病小鼠的白蛋白尿。