表观遗传修饰介导抑制SOSTDC1表达促进宫颈癌细胞的恶性生物学行为

目的:探究含硬化蛋白域蛋白1(SOSTDC1)对宫颈癌细胞恶性生物学行为的调控及其分子机制。方法:收集2020年8月至2022年5月间在福建省肿瘤医院活检或手术切除的53例宫颈癌组织和相应的癌旁组织标本,免疫组化法检测SOSTDC1蛋白在宫颈癌组织及相应癌旁组织中的表达,qPCR法检测正常宫颈细胞、宫颈癌细胞中SOSTDC1 mRNA表达;将SOSTDC1过表达慢病毒(OE-sostdc1)和对照空病毒(NC)感染宫颈癌细胞SiHa及CaSki,将其分为SiHa-OE-sostdc1、SiHa-NC、CaSki-OE-sostdc1、CaSki-NC组,采用WST-1法、细胞集落形成实验、Transwell实验和WB法检测转染各组SiHa及CaSki细胞的增殖、集落形成、迁移和侵袭能力和BMP、Wnt/β-catenin信号途径相关蛋白及上皮-间充质转化(EMT)相关蛋白的表达。用DNA甲基化酶抑制剂5-氮杂2'-脱氧胞苷(5'-Aza-CdR)处理宫颈癌细胞后采用qPCR和WB法检测SOSTDC1 m RNA及蛋白的表达变化,用甲基化特异性PCR(MSP)检测5例配对宫颈癌组织与癌旁组织中SOSTDC1基因启动子区甲基化水平,同时qPCR检测其SOSTDC1 mRNA水平。结果:与癌旁组织比较,SOSTDC1蛋白在宫颈癌组织中呈低表达(P<0.01),且与淋巴结转移与FIGO分期有关联(均P<0.05);与正常宫颈HUCEC细胞比较,SOSTDC1 mRNA在宫颈癌C33A、HeLa、SiHa、CaSki细胞中均呈低表达(均P<0.01)。过表达SOSTDC1显著抑制SiHa及CaSki细胞的增殖、迁移和侵袭能力(均P<0.05)。WB法结果检测显示,过表达SOSTDC1显著抑制SiHa及CaSki细胞中磷酸化Smad、Dvl2/3、β-catenin、VIM、N-cadherin、Snail蛋白的表达(均P<0.05),5'-Aza-CdR处理后的SiHa及CaSki细胞中SOSTDC1 mRNA和蛋白水平均显著增加(均P<0.05),MSP检测结果显示,相较于癌旁组织,宫颈癌组织中SOSTDC1基因启动子区呈高度甲基化,且SOSTDC1 mRNA水平降低(P<0.01)。结论:SOSTDC1在宫颈癌组织中呈低表达且与肿瘤的恶性进展关联,其表达下调与其基因启动子区高度甲基化有关,过表达SOSTDC1可能通过阻断BMP及Wnt/β-catenin信号通路从而抑制SiHa、CaSki细胞的增殖、侵袭和迁移能力。

SPOC domain-containing protein 1 regulates the proliferation and apoptosis of human spermatogonial stem cells through adenylate kinase 4

BACKGROUND Spermatogonial stem cells(SSCs)are the origin of male spermatogenesis,which can reconstruct germ cell lineage in mice.However,the application of SSCs for male fertility restoration is hindered due to the unclear mechanisms of proliferation and self-renewal in humans.AIM To investigate the role and mechanism of SPOC domain-containing protein 1(SPOCD1)in human SSC proliferation.METHODS We analyzed publicly available human testis single-cell RNA sequencing(RNAseq)data and found that SPOCD1 is predominantly expressed in SSCs in the early developmental stages.Small interfering RNA was applied to suppress SPOCD1 expression to detect the impacts of SPOCD1 inhibition on SSC proliferation and apoptosis.Subsequently,we explored the target genes of SPOCD1 using RNA-seq and confirmed their role by restoring the expression of the target genes.In addition,we examined SPOCD1 expression in some non-obstructive azoospermia(NOA)patients to explore the correlation between SPOCD1 and NOA.RESULTS The uniform manifold approximation and projection clustering and pseudotime analysis showed that SPOCD1 was highly expressed in the early stages of SSC,and immunohistological results showed that SPOCD1 was mainly localized in glial cell line-derived neurotrophic factor family receptor alpha-1 positive SSCs.SPOCD1 knockdown significantly inhibited cell proliferation and promoted apoptosis.RNA-seq results showed that SPOCD1 knockdown significantly downregulated genes such as adenylate kinase 4(AK4).Overexpression of AK4 in SPOCD1 knockdown cells partially reversed the phenotypic changes,indicating that AK4 is a functional target gene of SPOCD1.In addition,we found a significant downregulation of SPOCD1 expression in some NOA patients,suggesting that the downregulation of SPOCD1 may be relevant for NOA.CONCLUSION Our study broadens the understanding of human SSC fate determination and may offer new theories on the etiology of male infertility.

Combined TIM-3 and PD-1 blockade restrains hepatocellular carcinoma development by facilitating CD4+ and CD8+T cellmediated antitumor immune responses

BACKGROUND Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and T cell immunoglobulin and mucin domain-containing protein 3(TIM-3)are beneficial to the resumption of anti-tumor immunity response and hold extreme potential as efficient therapies for certain malignancies.However,ICIs with a single target exhibit poor overall response rate in hepatocellular carcinoma(HCC)patients due to the complex pathological mechanisms of HCC.AIM To investigate the effects of combined TIM-3 and PD-1 blockade on tumor development in an HCC mouse model,aiming to identify more effective immunotherapies and provide more treatment options for HCC patients.METHODS The levels of PD-1 and TIM-3 on CD4+and CD8+T cells from tumor tissues,ascites,and matched adjacent tissues from HCC patients were determined with flow cytometry.An HCC xenograft mouse model was established and treated with anti-TIM-3 monoclonal antibody(mAb)and/or anti-PD-1 mAb.Tumor growth in each group was measured.Hematoxylin and eosin staining and immunohistochemical staining were used to evaluate T cell infiltration in tumors.The percentage of CD4+and CD8+T cells in tissue samples from mice was tested with flow cytometry.The percentages of PD-1+CD8+,TIM-3+CD8+,and PD-1+TIM-3+CD8+T cells was accessed by flow cytometry.The levels of the cytokines including tumor necrosis factor alpha(TNF-α),interferon-γ(IFN-γ),interleukin(IL)-6,and IL-10 in tumor tissues were gauged with enzyme-linked immunosorbent assay kits.RESULTS We confirmed that PD-1 and TIM-3 expression was substantially upregulated in CD4+and CD8+T cells isolated from tumor tissues and ascites of HCC patients.TIM-3 mAb and PD-1 mAb treatment both reduced tumor volume and weight,while combined blockade had more substantial anti-tumor effects than individual treatment.Then we showed that combined therapy increased T cell infiltration into tumor tissues,and downregulated PD-1 and TIM-3 expression on CD8+T cells in tumor tissues.Moreover,combined treatment facilitated the production of T cell effector cytokines TNF-α and IFN-γ,and reduced the production of immunosuppressive cytokines IL-10 and IL-6 in tumor tissues.Thus,we implicated that combined blockade could ameliorate T cell exhaustion in HCC mouse model.CONCLUSION Combined TIM-3 and PD-1 blockade restrains HCC development by facilitating CD4+ and CD8+T cell-mediated antitumor immune responses.

巨噬细胞炎症蛋白-1α和骨硬化蛋白在多发性骨髓瘤中的表达及临床意义

目的研究多发性骨髓瘤(MM)患者骨髓上清液巨噬细胞炎症蛋白-1α(MIP-1α)和骨硬化蛋白(Sclerostin)的表达水平,探讨其在MM发病中的临床意义。方法采用酶链免疫法定量检测MIP-1α和Sclerostin的水平。结果 MM组中MIP-1α([152.44±96.29)pg/mL]和Sclerostin([0.53±0.24)pg/mL]水平明显高于对照者,两者比较差异有统计学意义(t值分别为5.69和4.70,P值分别为0.00和0.001);Ⅲ期MM患者的MIP-1α([172.91±102.41)pg/mL]和Sclerostin([0.61±0.22)pg/mL]的水平明显高于Ⅱ期患者(t值分别为3.70和2.87,P值分别为0.001和0.007);3级MBD患者的MIP-1α([191.41±110.82)pg/mL]和Sclerostin([0.68±0.19)pg/mL]明显高于非3级MBD患者(t值分别为3.01和5.69,P值分别为0.005和0.00);MM组中MIP-1α和Sclerostin表达呈正相关关系(r=0.498,P=0.002)。结论大部分MM患者骨髓MIP-1α和Sclerostin水平增高,其水平可反映体内瘤负荷及骨质破坏的程度,且两者具有明显相关性,联合检测可作为评估MM患者病变严重程度的一项指标。

血清中IL-7、PGLYRP-1、SOST的表达水平对类风湿性关节炎的诊断价值

目的探讨血清中白介素7(interleukin 7,IL-7)、肽聚糖识别蛋白1(peptidoglycan recognition protein 1,PGLYRP-1)、硬骨素(sclerostin,SOST)的表达水平对类风湿性关节炎的诊断价值。方法纳入2019年5月至2021年5月达州市中心医院收治的类风湿性关节炎患者82例为类风湿组、非类风湿性关节炎患者82例为非类风湿组、健康体检者82例为对照组。检测3组受试人员血清中IL-7, PGLYRP-1, SOST的表达量;用受试者工作特征曲线(receiver operating characteristic curve,ROC)的曲线下面积(area under curve,AUC)评估IL-7, PGLYRP-1, SOST检测在类风湿性关节炎中的诊断意义。结果类风湿组患者血清中的IL-7和SOST均低于非类风湿组和对照组(P<0.05),类风湿组患者血清中的PGLYRP-1高于非类风湿组患者和对照组(P<0.05)。IL-7、PGLYRP-1、SOST单向诊断类风湿性组患者与非类风湿组患者的灵敏度分别为90%、86%、98%,特异度分别为72%、89%、61%。血清中IL-7,PGLYRP-1, SOST联合对类风湿性关节炎的诊断的的灵敏度为91.46%,特异度为75.61%,准确度为80.89%。此外,非条件logistic逐步回归分析结果显示血清中IL-7, PGLYRP-1, SOST的表达水平均是患类风湿关节炎的重要危险因素。结论血清中IL-7、PGLYRP-1、SOST的表达水平可作为诊断类风湿性关节炎的潜在指标。

微RNA-21-5p靶向调控含硬化蛋白域蛋白1对甲状腺癌细胞生长的影响

目的:通过分析微RNA(microRNA,miR)-21-5p及其靶基因含硬化蛋白域蛋白1(recombinant sclerostin domain containing protein 1,SOSTDC1)在甲状腺癌中的作用,深入了解甲状腺癌转移的分子机制。方法:通过生物信息学分析和细胞验证筛选出miR-21-5p,通过miR-21-5p抑制剂转染甲状腺癌细胞系;采用MTT实验、流式细胞术和细胞划痕实验分别检测miR-21-5p抑制剂组和抑制剂对照组的甲状腺癌细胞增殖、凋亡和迁移的情况;采用荧光素酶报告实验验证miR-21-5p和SOSTDC1的靶向调控关系;采用蛋白质印迹法检测miR-21-5p抑制剂组和抑制剂对照组甲状腺癌细胞中SOSTDC1、下游磷脂酰肌醇3-激酶(phosphatidylinositol 3 kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)和丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)/细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)信号通路因子的表达水平及磷酸化水平。结果:MiR-21-5p在甲状腺癌细胞中显著上升,且与SOSTDC1呈负相关(r=-0.24,P<0.01);miR-21-5p抑制剂组甲状腺癌细胞增殖和迁移显著低于抑制剂对照组(均P<0.01),细胞凋亡率显著高于抑制剂对照组(P<0.01);荧光素酶报告实验表明miR-21-5p能够靶向调控SOSTDC1表达水平;甲状腺癌细胞中PI3K/Akt和MAPK/ERK信号通路检测显示miR-21-5p抑制剂组PI3K表达水平显著低于抑制剂对照组(P<0.01),Akt和ERK1/2水平无显著变化,但miR-21-5p抑制剂组Akt和ERK1/2磷酸化水平显著低于抑制剂对照组(均P<0.01)。结论:甲状腺癌细胞中miR-21-5p能够靶向抑制SOSTDC1的表达,影响PI3K/Akt和MAPK/ERK活性,从而抑制甲状腺癌细胞凋亡,促进细胞的增殖和迁移能力。

牙本质基质蛋白1在模拟失重骨丧失中的作用

目的检测模拟失重状态时牙本质基质蛋白1(dentin matrix protein 1,DMP1)和成骨抑制蛋白(sclerostin,SOST)在小鼠胫骨的表达,以及正常和DMP1基因缺失下SOST的表达,初步探讨DMP1在模拟失重下骨质丧失形成过程中的可能作用及机制。方法 1 1个月龄B6野生型小鼠随机分为悬尾组、对照组(n=5)。悬尾组悬吊1个月后与对照组胫骨石蜡切片进行HE及DMP1和SOST免疫组化染色,实时定量PCR检测Dmp1基因水平的表达。22个月龄B6Dmp1基因敲除鼠制备胫骨石蜡切片,进行SOST免疫组化染色。结果 HE染色显示悬尾组松质骨小梁较对照组稀疏,皮质骨变薄。免疫组化实验显示悬尾鼠DMP1和SOST在骨细胞细胞质和基质中的表达较对照组增高。高倍镜下DMP1和SOST阳性细胞数悬尾组显著高于对照组(P<0.05,P<0.01)。Dmp1实时定量PCR结果与免疫组化结果一致,悬尾组表达量为对照组的(2.8±0.4)倍,显著高于对照组(P<0.01)。SOST在Dmp1基因敲除鼠骨细胞中的表达也较正常增强。结论 DMP1在模拟失重小鼠胫骨中表达增强,提示无力学刺激这一特殊的机械信号可传递至骨细胞,导致其分泌DMP1的改变。SOST在骨形成中具有抑制作用,它在Dmp1基因敲除鼠及悬尾鼠胫骨中表达的增强提示SOST表达上调,导致骨形成减少可能是Dmp1基因敲除鼠和悬尾鼠骨质丧失的一个共同的重要因素。

补肾温阳止痛汤联合推拿治疗腰椎骨质增生的疗效及对血清DKK1、SOST、TXB2、6-Keto-PGFlα的影响

目的探讨补肾温阳止痛汤联合推拿治疗腰椎骨质增生的疗效及对血清Dickkopf相关蛋白1(DKK1)、硬骨素(SOST)、血栓素B2(TXB2)、6-酮前列腺素Flα(6-Keto-PGFlα)的影响。方法随机选取诊治的腰椎骨质增生患者80例,随机分为对照组与观察组,每组40例,对照组给口服塞来昔布胶囊及推拿治疗,观察组在以上治疗基础上联合补肾温阳止痛汤治疗,治疗4周。观察患者治疗前后中医证候变化,采用改良Oswestry功能障碍指数问卷(ODI)及腰椎生活量表(QLS-DSD)对患者腰功能及生活质量进行评价,观察血清DKK1、SOST、TXB2、6-Keto-PGFlα的变化。结果两组治疗后主证及次证积分均低于治疗前,且观察组低于对照组,差异有统计学意义(P<0.001);两组治疗后ODI及QLS-DSD评分均低于治疗前,且观察组低于对照组,差异有统计学意义(P<0.001);两组治疗后血清DKK1、SOST、6-Keto-PGFlα水平均高于治疗前,且观察组高于对照组,TXB2低于治疗前,且观察组低于对照组,差异有统计学意义(P<0.001)。结论补肾温阳止痛汤配合推拿治疗腰椎骨质增生可有效缓解症状,促进腰功能的恢复,提高生活质量,其作用可能与改善血清DKK1、SOST、TXB2、6-Keto-PGFlα水平有关。

脑血栓患者血清Dickkopf相关蛋白1、骨硬化蛋白表达水平及其与疾病严重程度的关系

目的探讨脑血栓患者血清中Dickkopf相关蛋白1(DKK-1)、骨硬化蛋白(SOST)水平与疾病严重程度的关系。方法选取110例脑血栓患者为脑血栓组,并根据严重程度分为轻、中、重度组;另选取同期一般资料相匹配的健康体检者110例为对照组。检测血清中DKK-1、SOST水平变化,并进行相关性分析;采用受试者工作特征(ROC)曲线分析DKK-1、SOST诊断重度脑血栓的效能。结果脑血栓组血清DKK-1、SOST水平高于对照组,差异有统计学意义(P<0.05);轻度组、中度组、重度组血清中DKK-1、SOST的水平呈显著升高趋势(P<0.05)。血清DKK-1与SOST水平存在正相关性(r=0.705,P<0.05),血清DKK-1、SOST水平均与脑血栓患者美国国立卫生研究院卒中量表(NIHSS)评分呈正相关性(r=0.625,r=0.674,P<0.05)。DKK-1、SOST诊断重度脑血栓的曲线下面积(AUC)分别为0.879(95%CI:0.808~0.950)、0.815(95%CI:0.710~0.919);DKK-1联合SOST诊断的AUC为0.911(95%CI:0.849~0.973)。结论脑血栓患者血清中DKK-1、SOST表达水平增高,且与疾病严重程度密切相关。DKK-1联合SOST对重度脑血栓有较高的诊断效能,对二者水平进行监测有助于评估患者疾病严重程度。

ATG16L1 and NOD2 polymorphisms enhance phagocytosis in monocytes of Crohn's disease patients

AIM: To investigate if the presence of relevant genetic polymorphisms has effect on the effectual clearance of bacteria by monocytes and granulocytes in patients with Crohn&#x02019;s disease (CD).

Sterile alpha motif and histidine-aspartic acid domain-containing protein 1 expression and its relationship with T cell activation in human immunodeficiency virus/acquired immune deficiency syndrome patients with lung-spleen Qi deficiency syndrome pattern

OBJECTIVE:To investigate the relationship between antiviral restriction factor Sterile Alpha Motif and Histidine-Aspartic acid domain-containing protein 1(SAMHD1)expression and T cell activation,furthermore,identifying objective indexes of lung-spleen Qi deficiency symptom pattern.METHODS:We assessed the profile of T lymphocyte subsets,characteristics of SAMHD1 and human leukocyte antigen DR(HLA-DR)expression in lungspleen Qi deficiency patients.At the same time,people living with human immunodeficiency virus/acquired immune deficiency syndrome(HIV/AIDS)(PLWHA)without obvious clinical symptoms and healthy donors in this area were used as controls.RESULTS:Immunohematologic indexes lower CD4 count,lower CD4/CD8 ratio and higher SAMHD1 level were found in lung-spleen Qi deficiency patients.Furthermore,we demonstrated a positive relationship between SAMHD1 and HLA-DR level as well as with interferon factor in lung-spleen Qi deficiency syndrome and patients without obvious clinical signs and symptoms groups.CONCLUSIONS:These data indicated the positive relationship between SAMHD1 and T cell activation which further elucidated the role of SAMHD1 in cellular immune response.Furthermore,combination of T lymphocyte subsets counts and SAMHD1 level may be used as clinical and biological reference basis for the differentiation and diagnosis of HIV/AIDS traditional Chinese medicine syndromes.

Effect of endothelial PAS domain protein 1 and hypoxia inducible factor 1~ on vascular endothelial growth factor expression in human pancreatic carcinoma

Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogenesis and vascular permeability of tumours, which promotes tumour growth and facilitates entry of cancer cells into blood circulation and metastasizing. This study examined whether HIF 1α and EPAS1 stimulated angiogenesis through activation of VEGF in human pancreatic carcinoma. Methods Specimens from pancreatic carcinoma and healthy parts of same pancreas were taken from 60 patients. Real time quantitative reverse transcription polymerase chain reaction estimated expression of HIF 1α, EPAS1, and VEGF mRNAs. Western blotting and immunohistochemical, streptavidin peroxidase method assessed expression of HIF 1α, EPAS1, and VEGF proteins. Microvessel density （MVD） was assessed. Results Highly significant increases in expression of EPAS1, VEGF, and MVD were found in pancreatic carcinoma tissue but not in normal pancreatic tissue： VEGF at mRNA and protein levels （t=17.32, P=-0.0001; t=98.41, P=0.0001）; EPAS1 protein level （t=22.51, P=0.0001）. Expression of HIF la was similar in pancreatic carcinoma and normal pancreatic tissues at both mRNA and protein levels. Significant correlations were observed between EPAS1 and VEGF （r=0.736, P=0.0041）, between VEGF and MVD （r=0.858, P=0.0001）, and between EPAS1 and MVD （r=0.641, P=0.0003）. No significant correlations were observed between HIF la and VEGF, or between HIF 1α and MVD. MVD and expression of EPAS1 and VEGF were significantly related with TNM staging, so was EPASI and VEGF with size of tumour. Conclusions EPAS1 and VEGF, but not HIFla, are overexpressed in pancreatic carcinoma. The expression of EPAS1 is correlated with that of VEGF and MVD. EPAS1 may be involved in the angiogenesis of pancreatic carcinoma by upregulating the expression of VEGE Targeting EPAS1 may be a new method of antiangiogenic tumour therapy for pancreatic carcinoma.

Structural insights of phosphorylated into the recognition FUNDC1 by LC3B in mitophagy

Mitophagy is an essential intracellular process that eliminates dysfunctional mitochondria and maintains cellular homeostasis. Mitophagy is regulated by the post-translational modification of mitophagy receptors. Fun14 domain-containing protein 1 （FUNDC1） was reported to be a new receptor for hypoxia-induced mitophagy in mammalian cells and interact with micro-tubule-associated protein light chain 3 beta （LC3B） through its LC3 interaction region （LIR）. Moreover, the phosphorylation modification of FUNDC1 affects its binding affinity for LC3B and regulates selective mitophagy. However, the structural basis of this regulation mechanism remains unclear. Here, we present the crystal structure of LC3B in complex with a FUNDCI LIR peptide phosphorylated at Ser17 （pS17）, demonstrating the key residues of LC3B for the specific recognition of the phosphorylated or dephosphorylated FUNDC1. Intriguingly, the side chain of LC3B Lys49 shifts remarkably and forms a hydrogen bond and electrostatic interaction with the phosphate group of FUNDC1 pS17. Alternatively, phosphorylated Tyr18 （PY18） and Ser13 （PS13） in FUNDC1 significantly obstruct their interaction with the hydrophobic pocket and Arg10 of LC3B, respectively. Structural observations are further validated by mutation and isothermal titration calorimetry （ITC） assays. Therefore, our structural and biochemical results reveal a working model for thespecific recognition of FUNDCI by LC3B and imply that the reversible phosphorylation modification of mitophagy receptors may be a switch for selective mitophagy.

Reduning plus ribavirin display synergistic activity against severe pneumonia induced by H1N1 influenza A virus in mice

OBJECTIVE:To investigate synergistic effect of Reduning(RDN)injection plus ribavirin against severe pneumonia induced by H1 N1 influenza A virus in mice.METHODS:We established a mouse model of severe pneumonia induced by influenza A virus by infecting Balb/c mice with CA07 virus.We randomly assigned the infected mice into four groups,and treated them with normal saline(NS group),RDN(injection,86.6 mg/kg),ribavirin(injection,66.6 mg/kg)or double Ribavirin plus RDN group,the same dosage as used in the single treatments)for 5 d.Lung index and lung pathology were recorded or calculated in terms of the curative effective.Cytokines,NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome related protein including caspase-associated recruitment domain(CARD)domain Apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1 and NOD-like receptor family,pyrin domain containing 3(NLRP3),and reactive oxygen species were simultaneously investigated.RESULTS:RDN plus ribavirin treatment,not RDN or ribavirin alone,provided a significant survival benefit to the influenza A virus-infected mice.The combination treatment protected the mice against severe influenza infection by attenuating the severe lung injury.The combined treatment also reduced the viral titers in mouse lungs and lung index,downregulated their immunocytokine levels,including IL-1βand IL-18,and down regulated the NLRP3,especially the transcription and translation of caspase-1.Meanwhile NS group had significantly higher reactive oxygen species(ROS)expression which could was dramatically reduced by the treatment of RDN plus ribavirin.CONCLUSION:Our study showed that RDN combined with ribavirin could protect the mice,and reduce the lung immunopathologic damage caused by severe influenza pneumonia.The mechanism could be that it reduced ROS produce and inhibited NLRP3 inflammasome activation so that mainly lower the downstream inflammatory cytokines IL-1βand IL-18.

Interaction of the major inflammatory bowel disease susceptibility alleles in Crohn’s disease patients

AIM:To investigate the interaction of interleukin-23 receptor(IL23R)(rs1004819 and rs2201841),autophagy-related 16-like 1(ATG16L1)(rs2241880), caspase recruitment domain-containing protein 15 (CARD15)genes,and IBD5 locus in Crohn's disease(CD) patients. METHODS:A total of 315 unrelated subjects with CD and 314 healthy controls were genotyped.Interactions and specific genotype combinations of a total of eight variants were tested.The variants of IBD5locus(IGR2198a_1 rs11739135 and IGR2096a_1 rs12521868),CARD15(R702W rs2066845 and L1007fs rs2066847),ATG16L1(rs2241880)and IL23R (rs1004819,rs2201841)genes were genotyped by PCR-RFLP,the G908R(rs2066844)in CARD15 was determined by direct sequencing. RESULTS:The association of ATG16L1 T300A with CD was confirmed[P=0.004,odds ratio(OR)=1.69, 95%CI:1.19-2.41],and both IL23R variants were found to represent significant risk for the disease(P= 0.008,OR=2.05,95%CI:1.20-3.50 for rs1004819 AA;P<0.001,OR=2.97,95%CI:1.65-5.33 for rs2201841 CC).Logistic regression analysis of pairwise interaction of the inflammatory bowel disease (IBD)loci indicated that IL23R,ATG16L1,CARD15 and IBD5(IGR2198a_1)contribute independently to disease risk.We also analysed the specific combina- tions by pair of individual ATG16L1,IL23R rs1004819, rs2201841,IGR2198a_1,IGR2096a_1 and CARD15 genotypes for disease risk influence.In almost all cases,the combined risk of susceptibility pairs was higher in patients carrying two different risk-associated gene variants together than individuals with just one polymorphism.The highest OR was found for IL23R rs2201841 homozygous genotype with combination of positive CARD15 status(P<0.001,OR=9.15,95% CI:2.05-40.74). CONCLUSION:The present study suggests a cumulative effect of individual IBD susceptibility loci.

Targeting neuronal mitophagy in ischemic stroke:an update

Cerebral ischemia is a neurological disorder associated with complex pathological mechanisms,including autophagic degradation of neuronal mitochondria,or termed mitophagy,following ischemic events.Despite being well-documented,the cellular and molecular mechanisms under-lying the regulation of neuronal mitophagy remain unknown.So far,the evidence suggests neuronal autophagy and mitophagy are separately regulated in ischemic neurons,the latter being more likely activated by reperfusional injury.Specifically,given the polarized morphology of neurons,mitophagy is regulated by different neuronal compartments,with axonal mitochondria being degraded by autophagy in the cell body following ischemia-reperfusion insult.A variety of molecules have been associated with neuronal adaptation to ischemia,including PTEN-induced kinase 1,Parkin,BCL2 and adenovirus E1B 19-kDa-interacting protein 3(Bnip3),Bnip3-like(Bnip3l)and FUN14 domain-containing 1.Moreover,it is still controversial whether mitophagy protects against or instead aggravates ischemic brain injury.Here,we review recent studies on this topic and provide an updated overview of the role and regulation of mitophagy during ischemic events.

Reverse effect of Semaphorin-3F on rituximab resistance in diffuse large B-cell lymphoma via the Hippo pathway

Background:Exploring the underlying mechanism of rituximab resistance is critical to improve the outcomes of patients with diffuse large B-cell lymphoma(DLBCL).Here,we tried to identify the effects of the axon guidance factor semaphorin-3F(SEMA3F)on rituximab resistance as well as its therapeutic value in DLBCL.Methods:The effects of SEMA3F on the treatment response to rituximab were investigated by gain-or loss-of-function experiments.The role of the Hippo pathway in SEMA3F-mediated activity was explored.A xenograft mouse model generated by SEMA3F knockdown in cells was used to evaluate rituximab sensitivity and combined therapeutic effects.The prognostic value of SEMA3F and TAZ(WW domain-containing transcription regulator protein 1)was examined in the Gene Expression Omnibus(GEO)database and human DLBCL specimens.Results:We found that loss of SEMA3F was related to a poor prognosis in patients who received rituximab-based immunochemotherapy instead of chemotherapy regimen.Knockdown of SEMA3F significantly repressed the expression of CD20 and reduced the proapoptotic activity and complement-dependent cytotoxicity(CDC)activity induced by rituximab.We further demonstrated that the Hippo pathway was involved in the SEMA3F-mediated regulation of CD20.Knockdown of SEMA3F expression induced the nuclear accumulation of TAZ and inhibited CD20 transcriptional levels via direct binding of the transcription factor TEAD2 and the CD20 promoter.Moreover,in patients with DLBCL,SEMA3F expression was negatively correlated with TAZ,and patients with SEMA3F^(low)TAZ^(high)had a limited benefit from a rituximab-based strategy.Specifically,treatment of DLBCL cells with rituximab and a YAP/TAZ inhibitor showed promising therapeutic effects in vitro and in vivo.Conclusion:Our study thus defined a previously unknown mechanism of SEMA3F-mediated rituximab resistance through TAZ activation in DLBCL and identified potential therapeutic targets in patients.

Cystathionine-γ-lyase ameliorates the histone demethylase JMJD3-mediated autoimmune response in rheumatoid arthritis

Cystathionine-γ-lyase(CSE),an enzyme associated with hydrogen sulfide(H2S)production,is an important endogenous regulator of inflammation.Jumonji domain-containing protein 3(JMJD3)is implicated in the immune response and inflammation.Here,we investigated the potential contribution of JMJD3 to endogenous CSE-mediated inflammation in rheumatoid arthritis(RA).Upregulated CSE and JMJD3 were identified in synovial fibroblasts(SFs)from RA patients as well as in the joints of arthritic mice.Knocking down CSE augmented inflammation in IL-1β-induced SFs by increasing JMJD3 expression.In addition,CSE−/−mice with collagen-induced arthritis(CIA)developed severe joint inflammation and bone erosion.Conversely,overexpressing CSE inhibited JMJD3 expression by the transcription factor Sp-1 and was accompanied by reduced inflammation in IL-1β-treated SFs.Furthermore,JMJD3 silencing or the administration of the JMJD3 inhibitor GSK-J4 significantly decreased the inflammatory response in IL-1β-treated SFs,mainly by controlling the methylation status of H3K27me3 at the promoter of its target genes.GSK-J4 markedly attenuated the severity of arthritis in CIA mice.In conclusion,suppressing JMJD3 expression by the transcription factor Sp-1 is likely responsible for the ability of CSE to negatively modulate the inflammatory response and reduce the progression of RA.