Transforming liquid flow fuel cells to controllable reactors for highlyefficient oxidation of 5-hydroxymethylfurfural to 2, 5-furandicarboxylic acid at low temperature

Highly-efficient oxidation of 5-hydroxymethylfurtural(HMF) to 2,5-furandicarboxylic acid(FDCA) at low temperature with air as the oxidant is still challenging.Herein,inspired by the respirato ry electron transport chain(ETC) of living cells mediated by electron carriers,we constructed artificial ETCs and transformed liquid flow fuel cells(LFFCs) to flexible reactors for efficient oxidation of HMF to produce FDCA under mild conditions.This LFFC reactor employed an electrodeposition modified nickel foam as an anode to promote HMF oxidation and(VO_(2))_(2)SO_(4) as a cathode electron carrier to facilitate the electron transfer to air.The reaction rate could be easily controlled by selecting the anode catalyst,adjusting the external loading and changing the cathodic electron carrier or oxidants.A maximal power density of 44.9 mW cm^(-2) at room temperature was achieved,while for FDCA production,short-circuit condition was preferred to achieve quick transfer of electrons.For a single batch operation with 0.1 M initial HMF,FDCA yield reached 97.1%.By fed-batch operation,FDCA concentration reached 144.5 g L^(-1) with a total yield of 96%.Ni^(2+)/Ni^(3+) redox couple was the active species mediating the electron transfer,while both experimental and DFT calculation results indicated that HMFCA pathway was the preferred reaction mechanism.

Perilipin 5 regulates hepatic stellate cell activation and high-fat diet-induced non-alcoholic fatty liver disease

Background:Nonalcoholic fatty liver disease(NAFLD)is one of the most common chronic liver diseases globally.Hepatic stellate cells(HSCs)are the major effector cells of liver fibrosis.HSCs contain abundant lipid droplets(LDs)in their cytoplasm during quiescence.Perilipin 5(PLIN 5)is a LD surface-associated protein that plays a crucial role in lipid homeostasis.However,little is known about the role of PLIN 5 in HSC activation.Methods:PLIN 5 was overexpressed in HSCs of Sprague–Dawley rats by lentivirus transfection.At the same time,PLIN 5 gene knockout mice were constructed and fed with a high-fat diet(HFD)for 20 weeks to study the role of PLIN 5 in NAFLD.The corresponding reagent kits were used to measure TG,GSH,Caspase 3 activity,ATP level,and mitochondrial DNA copy number.Metabolomic analysis of mice liver tissue metabolism was performed based on UPLC-MS/MS.AMPK,mitochondrial function,cell proliferation,and apoptosis-related genes and proteins were detected by western blotting and qPCR.Results:Overexpression of PLIN 5 in activated HSCs led to a decrease in ATP levels in mitochondria,inhibition of cell proliferation,and a significant increase in cell apoptosis through AMPK activation.In addition,compared with the HFD-fed C57BL/6J mice,PLIN 5 knockout mice fed with HFD showed reduced liver fat deposition,decreased LD abundance and size,and reduced liver fibrosis.Conclusion:These findings highlight the unique regulatory role of PLIN 5 in HSCs and the role of PLIN 5 in the fibrosis process of NAFLD.

Exosome-Transmitted miR-224-5p Promotes Colorectal Cancer Cell Proliferation via Targeting ULK2 in p53-Dependent Manner

Objective To investigate the role and molecular mechanism of exosomal miR-224-5p in colorectal cancer(CRC).Methods The miR-224-5p expression in CRC patient tissues and cell-derived exosomes was measured by laser capture microdissection and qRT-PCR,respectively.Dual-luciferase reporter gene assay was used to determine the target gene of miR-224-5p.The protein expressions of p53 and unc-51 like kinase 2(ULK2)in CRC cells were detected by western blot.Flow cytometry was used to detect cell cycle and apoptosis.Cell proliferation was measured by CCK8 and EdU assay.Results The miR-224-5p expression was upregulated in CRC tissues and increased progressively with the rise of CRC stage.CRC cells secreted extracellular miR-224-5p mainly in an exosome-dependent manner,and then miR-224-5p could be transferred to surrounding tumor cells to regulate cell proliferation in the form of autocrine or paracrine.Moreover,ULK2 was characterized as a direct target of miR-224-5p and was downregulated in CRC tissues.Interestingly,ULK2 inhibited CRC cell proliferation in a p53-dependent manner.Furthermore,exosome-derived miR-224-5p partially reversed the proliferation regulation of ULK2 on CRC cells.Conclusion Our findings demonstrate that exosome-transmitted miR-224-5p promotes p53-dependent cell proliferation by targeting ULK2 in CRC,which may offer promising targets for CRC prevention and therapy.

Differentiation and immunosuppressive function of CD19~+CD24~(hi)CD27~+ regulatory B cells are regulated through the miR-29a-3p/NFAT5 pathway

Background: Regulatory B cells(Bregs) is an indispensable element in inducing immune tolerance after liver transplantation. As one of the microRNAs(miRNAs), mi R-29a-3p also inhibits translation by degrading the target mRNA, and yet the relationship between Bregs and mi R-29a-3p has not yet been fully explored. This study aimed to investigate the impact of miR-29a-3p on the regulation of differentiation and immunosuppressive functions of memory Bregs(m Bregs) and ultimately provide potentially effective therapies in inducing immune tolerance after liver transplantation. Methods: Flow cytometry was employed to determine the levels of Bregs in peripheral blood mononuclear cells. TaqMan low-density array miRNA assays were used to identify the expression of different miRNAs, electroporation transfection was used to induce mi R-29a-3p overexpression and knockdown, and dual luciferase reporter assay was used to verify the target gene of miR-29a-3p. Results: In patients experiencing acute rejection after liver transplantation, the proportions and immunosuppressive function of m Bregs in the circulating blood were significantly impaired. mi R-29a-3p was found to be a regulator of m Bregs differentiation. Inhibition of miR-29a-3p, which targeted nuclear factor of activated T cells 5(NFAT5), resulted in a conspicuous boost in the differentiation and immunosuppressive function of m Bregs. The inhibition of mi R-29a-3p in CD19~+ B cells was capable of raising the expression levels of NFAT5, thereby promoting B cells to differentiate into m Bregs. In addition, the observed enhancement of differentiation and immunosuppressive function of m Bregs upon mi R-29a-3p inhibition was abolished by the knockdown of NFAT5 in B cells. Conclusions: mi R-29a-3p was found to be a crucial regulator for m Bregs differentiation and immunosuppressive function. Silencing mi R-29a-3p could be a potentially effective therapeutic strategy for inducing immune tolerance after liver transplantation.

Small cell lung carcinoma with KIF5B-RET fusion partially responded to the 4^(th)-line therapy with anlotinib:A case report

BACKGROUND Small cell lung cancer(SCLC)exhibits a pronounced tendency for metastasis and relapse,and the acquisition of resistance to chemotherapy and radiotherapy,leading to complexity in treatment outcomes.It is crucial to tackle these challenges by advancing targeted therapeutic approaches in ongoing research endeavors.Variant RET fusions have been reported in several solid tumors,but are rarely reported in SCLC.CASE SUMMARY We present the first case of a KIF5B-RET fusion in a 65-year-old male patient with SCLC.To date,the patient has received the 4th line chemotherapy with anlotinib for one year and has shown a sustained favorable partial response.According to the results of next generation sequencing,this SCLC patient harbors the KIF5BRET fusion,suggesting that RET fusion could serve as a promising molecular target for SCLC treatment.Next-generation sequencing(NGS)plays a critical rolein comprehensively assessing the genotype and phenotype of cancer.CONCLUSION NGS can provide SCLC patients with personalized and targeted therapy options,thereby improving their likelihood of survival.

Ghrelin regulates insulin resistance by targeting insulin-like growth factor-1 receptor via miR-455-5p in hepatic cells

Objective: To explore the mechanism by which ghrelin regulates insulin sensitivity through modulation of miR-455-5p in hepatic cells. Methods: HepG2 cells were treated with or without DAG (1 μM). Glucose consumption, intracellular glycogen content, phosphorylation of PI3K and Akt stimulated by insulin, expression of miR-455-5p, as well as IGF-1R protein level were analyzed. In addition, bioinformatic analysis, dual luciferase reporter assay, miR- 455-5p mimic or inhibitor treatment was conducted to investigate the molecular mechanisms. Results: High glucose treatment upregulated miR-455-5p expression but reduced glucose consumption and glycogen content. DAG reversed the effect of high glucose on glucose metabolism, increased protein level of IGF-1R and phosphorylation of PI3K/Akt stimulated by insulin, as well as downregulated miR-455-5p expression. Bioinformatic analysis indicated IGF-1R was the target of miR-455-5p. Dual luciferase reporter assay, as well as transfection with miR-455-5p mimic/inhibitor confirmed that DAG activated IGF-1R/PI3K/Akt signaling via inhibiting miR-455-5p. Conclusion: DAG improves insulin resistance via miR-455-5p- mediated activation of IGF-1R/PI3K/Akt system, suggesting that suppression of miR-455-5p or activation of DAG may be potential targets for T2DM therapy.

Inhibitory Effect of Flavonoid Glycosides from Chlorophytum comosum on Nasopharyngeal Carcinoma 5-8F Cells and Its Mechanism

[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavonoid glycosides were isolated and purified from the ethanol alcoholic extract of the roots of Liliaceae plant Chlorophytum comosum by silica gel column chromatography,macroporous resin column chromatography,Sephadex LH-20,and reverse column chromatography(ODS).The inhibitory activity of flavonoid glycosides on human nasopharyngeal carcinoma cells was analyzed by CCK-8 method,and the potential mechanism was preliminarily analyzed by molecular docking.[Results]Two flavonoid glycosides were identified as isovitexin 2″-0-rhamnoside and 7-2″-di-O-β-glucopyranosylisovitexin.Two flavonoid glycosides showed promising inhibitory effect on human nasopharyngeal carcinoma cell line 5-8F,with IC_(50) values of 24.8 and 27.5μmol/L,respectively.Molecular docking results showed that the potential targets of two flavonoid glycosides include CyclinD1,Bcl-2β-Catenin,ILK,TGF-β,in addition,two glycosides showed higher predicted binding affinity towards CyclinD1,which verifies the cytotoxicity of the two compounds on human nasopharyngeal carcinoma cell line 5-8F in vitro.[Conclusions]Two flavonoid glycosides are the active molecules in Chlorophytum comosum that can inhibit the proliferation of human nasopharyngeal carcinoma cells,and have the potential to be used in the research and development of anti nasopharyngeal carcinoma drugs.

Gamma-glutamyl transferase 5 overexpression in cerebrovascular endothelial cells improves brain pathology,cognition,and behavior in APP/PS1 mice

In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of Alzheimer’s disease remains unclear.This study aimed to explore the effect of GGT5 on cognitive function and brain pathology in an APP/PS1 mouse model of Alzheimer’s disease,as well as the underlying mechanism.We observed a significant reduction in GGT5 expression in two in vitro models of Alzheimer’s disease(Aβ_(1-42)-treated hCMEC/D3 and bEnd.3 cells),as well as in the APP/PS1 mouse model.Additionally,injection of APP/PS1 mice with an adeno-associated virus encoding GGT5 enhanced hippocampal synaptic plasticity and mitigated cognitive deficits.Interestingly,increasing GGT5 expression in cerebrovascular endothelial cells reduced levels of both soluble and insoluble amyloid-βin the brains of APP/PS1 mice.This effect may be attributable to inhibition of the expression ofβ-site APP cleaving enzyme 1,which is mediated by nuclear factor-kappa B.Our findings demonstrate that GGT5 expression in cerebrovascular endothelial cells is inversely associated with Alzheimer’s disease pathogenesis,and that GGT5 upregulation mitigates cognitive deficits in APP/PS1 mice.These findings suggest that GGT5 expression in cerebrovascular endothelial cells is a potential therapeutic target and biomarker for Alzheimer’s disease.

Mechanism of stilbene glycosides on apoptosis of SH-SY5Y cells via regulating PI3K/AKT signaling pathway

Objective:To investigate the effects of stilbene glycoside(TSG)on okadaic acid-induced apoptosis in human neuroblastoma cells(SH-SY5Y)via the PI3K/AKT pathway.Methods:The optimal concentration of OA was screened by CCK-8 assay,and SH-SY5Y cells were divided into control group,model group,TSG group,LY294002 group and LY294002+TSG group.The proliferation and apoptosis in each group were detected by CCK-8 and TUNEL assays;Western blotting method and real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of PI3K,P-PI3K(Y607),AKT,P-AKT(Ser473),Bcl-2 and Bax proteins.The relative protein expression was represented by P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax gray ratio.Results:CCK-8 screened the optimal concentration of OA as 40 nmol/L.Compared with the control group,the model group increased relative cell viability,decreased apoptosis rate,the pathway and apoptotic proteins expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax were decreased,and the mRNA expression levels of PI3K,AKT and Bcl-2 were decreased.Bax mRNA expression level increased(P<0.05);Compared with model group,TSG group increased relative cell viability,decreased apoptosis rate,increased protein expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT,Bcl-2/Bax,and increased mRNA expression levels of PI3K,AKT,and Bcl-2.Bax mRNA expression decreased(P<0.05),LY294002 group decreased relative cell viability,increased apoptosis rate,P-PI3K(Y607)/PI3K protein expression levels were significantly decreased(P<0.05),P-AKT(Ser473)/AKT and Bcl-2/Bax protein expression levels were significantly decreased,but there was no statistical significance,PI3K,AKT and Bcl-2 mRNA expression levels were decreased,and Bax mRNA expression levels were increased(all P<0.05);Compared with LY294002 group,LY294002+TSG group increased relative cell viability,decreased apoptosis rate,and the protein expression levels of P-PI3K(Y607)/PI3K,P-AKT(Ser473)/AKT and Bcl-2/Bax were increased.The mRNA expression levels of PI3K,AKT,Bcl-2 were increased,Bax was decreased(all P<0.05).Conclusion:Stilbene glycoside may alleviate okadaic acid-induced apoptosis in SH-SY5Y cells by interfering with the PI3K/AKT signaling pathway,which in turn regulates the expression of apoptotic factors such as Bcl-2 and Bax.

非小细胞肺癌组织中lncRNA GAS5、LHPP表达与上皮间质化相关性及临床意义

目的探讨非小细胞肺癌(NSCLC)患者癌组织中长链非编码核糖核酸生长抑制特异性基因5(lncRNA GAS5)、磷酸赖氨酸磷酸组氨酸无机焦磷酸盐磷酸酶(LHPP)表达与上皮间质化(EMT)相关性及临床意义。方法收集2018年6月至2020年1月在河北北方学院附属第一医院行手术切除的NSCLC 90例患者的癌组织及癌旁组织,采用实时荧光定量聚合酶链反应检测lncRNA GAS5、LHPP和EMT相关蛋白[E-钙黏蛋白(E-Cad)、N-钙黏蛋白(N-Cad)和波形蛋白(VIM)]表达。分析NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA与临床病理特征的关系,并通过Pearson相关性分析NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA与EMT相关蛋白表达的相关性。采用Kaplan-Meier法绘制不同lncRNA GAS5、LHPP mRNA表达的NSCLC患者生存曲线,多因素Cox回归分析NSCLC患者预后的影响因素。结果NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA、E-Cad mRNA表达低于癌旁组织,N-Cad mRNA、VIM mRNA表达高于癌旁组织,差异有统计学意义(P<0.05)。Pearson相关性分析显示,NSCLC患者癌组织中lncRNA GAS5与E-Cad mRNA表达呈正相关(r=0.724,P<0.001),与N-Cad mRNA、VIM mRNA表达呈负相关(r=-0.699、-0.689,P<0.001);lncRNA GAS5与LHPP mRNA表达呈正相关(r=0.651,P<0.001)。不同分化程度、肿瘤TNM分期、淋巴结转移的NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA表达比较差异有统计学意义(P<0.05)。Kaplan-Meier生存曲线分析显示,lncRNA GAS5高表达组的3年总生存率[68.18%(30/44)]高于lncRNA GAS5低表达组的3年总生存率[36.96%(17/46)];LHPP mRNA高表达组的3年总生存率[67.39%(31/46)]高于LHPP mRNA低表达组的3年总生存率[36.36%(16/44)],差异有统计学意义(χ^(2)=10.274、10.322,P<0.05)。低分化、肿瘤TNM分期Ⅲ期、有淋巴结转移为NSCLC患者死亡的独立危险因素,lncRNA GAS5≥1.32、LHPP mRNA≥1.12为独立保护因素(P<0.05)。结论NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA低表达,与EMT相关蛋白表达、分化程度、肿瘤TNM分期、淋巴结转移和预后有关,可能成为NSCLC诊治的新靶点。

拮抗CC趋化因子受体5信号诱导肿瘤细胞凋亡并调节肿瘤微环境抑制肿瘤生长

目的探索拮抗CC趋化因子受体5(CCR5)信号通路对肿瘤生长和肿瘤微环境的影响。方法采用CCK8细胞毒试验研究CCR5选择性拮抗剂Maraviroc体外对小鼠Lewis肺腺癌细胞增殖的影响,并运用流式细胞术和RT-PCR检测肿瘤细胞凋亡和Caspase 8基因的表达。然后采用免疫荧光组织化学染色法研究了Maraviroc对小鼠体内肿瘤生长和肿瘤微环境中CD4^(+)和CD8^(+)以及Foxp3^(+)细胞比例的影响。结果拮抗CCR5信号在体内外均能够抑制癌细胞的生长。体外研究发现:CCR5拮抗剂可通过增强凋亡基因Caspase 8表达而诱导肿瘤细胞凋亡。在小鼠体内,CCR5拮抗剂可明显增加肿瘤微环境中CD4^(+)和CD8^(+)细胞的浸润而减少Foxp3^(+)细胞的浸润。结论拮抗CCR5信号可能通过诱导肿瘤细胞凋亡,逆转免疫抑制性肿瘤微环境而抑制肿瘤生长。

SLC6A9对结直肠癌细胞生长和对5-FU药物敏感性的影响

目的:研究溶质载体家族6成员9(solute carrier family 6 member 9,SLC6A9)表达对结直肠癌细胞增殖、迁移和5-氟尿嘧啶(5-fluorouracil,5-FU)药物敏感性的影响。方法:TCGA数据库分析、实时荧光定量PCR和Western blot分析检测SLC6A9在结肠癌组织、正常结肠细胞系(NCM460)和结直肠癌细胞系(SW620、HCT116、HT29、Lovo和SW480)中的表达。将SCL6A9过表达质粒及阴性对照(SLC6A9 OE、Vector)转染HT29细胞,将SCL6A9小干扰RNA及阴性对照(SLC6A9 siRNA1#、siRNA2#和Scramble)转染SW620细胞。划痕愈合实验和Transwell实验检测各组细胞的迁移、侵袭能力。Western blot和细胞免疫荧光检测EMT相关蛋白E-cadherin、Vimentin的表达水平。利用CCK-8法和构建裸鼠移植瘤模型检测SLC6A9过表达对结直肠癌细胞5-FU药物敏感性的影响。结果:与正常结肠组织和NCM460细胞相比,SLC6A9在结肠癌组织和结直肠癌细胞系中低表达(均P<0.05)。SLC6A9过表达引起E-cadherin蛋白表达增加,Vimentin蛋白水平降低,抑制结直肠癌细胞的迁移、侵袭(P<0.05)。SLC6A9低表达引起E-cadherin蛋白表达降低,Vimentin蛋白水平增加,促进结直肠癌细胞的迁移、侵袭能力(P<0.05)。SLC6A9过表达提高了5-FU的药物敏感性,并使肿瘤生长缓慢,质量减轻(P<0.05)。而SLC6A9低表达降低了5-FU的药物敏感性(P<0.05)。结论:SLC6A9过表达能够抑制结直肠癌细胞的迁移、侵袭和EMT进程,并增强5-FU对结直肠癌细胞的药物敏感性。

CXCL5通过诱导血管钙化参与颈动脉斑块的形成

背景:CXC基序趋化因子5(CXC-motif chemokine 5,CXCL5)为上皮细胞衍生的中性粒细胞激活肽,研究发现其可能参与动脉病变。然而,CXCL5在血管钙化中的作用未见报道。目的:探讨CXCL5在颈动脉粥样硬化的血管钙化中的作用。方法:①细胞实验:将小鼠血管平滑肌细胞分成以下各组:成骨培养基组,Vector组(空白质粒转染到细胞中),CXCL5组(CXCL5质粒转染到细胞中),si-NC组(CXCL5阴性对照siRNA转染到细胞中),si-CXCL5组(CXCL5 siRNA转染到细胞中),Vector+LY2157299组和CXCL5+LY2157299组(细胞转染24 h后,将转化生长因子β受体1激酶抑制剂LY2157299加入细胞中)。进行茜素红染色、碱性磷酸酶染色和钙含量测定以评估血管平滑肌细胞成骨分化水平。②动物实验:48只ApoE-/-小鼠随机分成4组:Con+si-NC组、Con+si-CXCL5组、CAS+si-NC组和CAS+si-CXCL5组,前2组不造模,尾静脉注射si-NC或si-CXCL5慢病毒;后2组制备颈动脉粥样硬化模型,尾静脉注射si-NC或si-CXCL5慢病毒。采用Von Kossa染色和免疫组织化学染色评估小鼠颈动脉血管钙化以及CXCL5、转化生长因子β受体1表达情况。结果与结论:①CXCL5组细胞Runt相关转录因子2蛋白水平上调、α-平滑肌肌动蛋白水平下调,si-CXCL5组中的发现与其相反;CXCL5过表达上调了转化生长因子β受体1水平,而CXCL5敲低抑制了转化生长因子β受体1水平。②与Vector组相比,CXCL5组细胞茜素红染色的强度、碱性磷酸酶活性和钙含量显著增加(P<0.05);与si-NC组相比,si-CXCL5组上述2项指标显著降低(P<0.05);当用LY2157299抑制转化生长因子β受体1表达时,CXCL5对平滑肌细胞的成骨转化作用减弱。③与Con+si-NC组相比,CAS+si-NC组大鼠颈动脉中CXCL5蛋白表达和血管钙化面积显著增加(P<0.05);与CAS+si-NC组相比,CAS+si-CXCL5组颈动脉中上述2项指标显著降低(P<0.05)。④与Con+si-NC组相比,CAS+si-NC组大鼠颈动脉中Runt相关转录因子2蛋白表达显著增加(P<0.05)和α-平滑肌肌动蛋白表达显著降低(P<0.05);与CAS+si-NC组相比,CAS+si-CXCL5组颈动脉中上述2项指标呈相反变化(P<0.05)。⑤结果说明,CXCL5通过激活转化生长因子β受体1通路诱导血管平滑肌细胞成骨样转化,抑制CXCL5表达对于改善颈动脉粥样硬化小鼠颈动脉血管钙化是有效的。

miR-592靶向调控PRDM5影响肾细胞癌侵袭转移及自噬的机制研究

目的探究miR-592靶向调控PR结构域锌指蛋白5(PR domain zinc finger protein 5,PRDM5)影响肾细胞癌(renal cell carcinoma,RCC)侵袭转移及自噬的机制。方法RT-qPCR、蛋白质印迹检测RCC组织和细胞中miR-592、PRDM5 mRNA和蛋白水平。A498、786-O细胞分为anti-miR-NC组、anti-miR-592组、anti-miR-592+si-NC组、anti-miR-592+si-PRDM5组。RNA免疫共沉淀(RIP)检测miR-592和PRDM5的结合;CCK-8、Transwell实验检测细胞增殖、迁移和侵袭能力;透射电镜观察自噬小体形成;蛋白质印迹检测细胞PRDM5、基质金属蛋白酶(MMP)-2、MMP-9、Beclin1、微管相关蛋白1轻链3(LC3)Ⅱ/Ⅰ水平。建立移植瘤裸鼠模型,分析敲低miR-592表达对移植瘤生长的影响。结果RCC组织和细胞中PRDM5 mRNA和蛋白水平降低,miR-592水平增加(P<0.05)。敲低miR-592表达后,细胞A_(450 nm)、迁移和侵袭细胞数、MMP-2、MMP-9水平降低,自噬小体数量、Beclin1、LC3Ⅱ/Ⅰ水平增加(P<0.05)。敲低PRDM5表达能部分逆转敲低miR-592对细胞增殖、侵袭及自噬的影响(P<0.05)。敲低miR-592表达能够抑制裸鼠移植瘤生长(P<0.05)。结论miR-592通过靶向抑制PRDM5表达,促进RCC细胞增殖、侵袭转移能力,并抑制自噬能力。

DCLK1和ITGA5在非小细胞肺癌患者中的表达及其与临床病理特征及预后的关系研究

目的 检测非小细胞肺癌(non-small cell lung cancer,NSCLC)患者血清双皮质素样激酶1(doublecortin-like kinase 1,DCLK1)和整合素α5(integrin alpha5,ITGA5)水平并讨论二者与临床病理特征及预后的关系。方法 选取2017年6月1日至2020年7月1日在我院收治的116例NSCLC患者(NSCLC组)作为研究对象,收集同期100例健康受试者作为对照组,采用酶联免疫吸附法检测NSCLC组和对照组受试者血清中DCLK1和ITGA5的水平;根据预后情况将患者分为生存组和死亡组,ROC曲线分析血清DCLK1、ITGA5的水平对NSCLC患者3年发生死亡的预测价值,多因素Cox回归分析影响NSCLC患者发生死亡的因素。结果 NSCLC组患者血清DCLK1、ITGA5的水平高于对照组(P<0.05);不同吸烟史、TNM分期、分化程度、肿瘤直径以及淋巴结转移的NSCLC患者血清DCLK1、ITGA5的水平比较,差异有统计学意义(P<0.05);死亡组患者血清DCLK1、ITGA5的水平均高于生存组(P<0.05);血清DCLK1、ITGA5单独及联合预测患者发生死亡的AUC分别为0.895(95%CI=0.824~0.944)、0.828(95%CI=0.747~0.892)、0.926(95%CI=0.862~0.966);DCLK1、ITGA5及淋巴结转移是影响NSCLC患者发生死亡的独立危险因素(P<0.05)。结论 NSCLC患者血清DCLK1、ITGA5的水平异常升高,二者的表达水平与NSCLC患者临床病理特征和不良预后有关。

The circ_0002538/miR-138-5p/plasmolipin axis regulates Schwann cell migration and myelination in diabetic peripheral neuropathy

Circular RNAs(circRNAs)play a vital role in diabetic peripheral neuropathy.However,their expression and function in Schwann cells in individuals with diabetic peripheral neuropathy remain poorly understood.Here,we performed protein profiling and circRNA sequencing of sural nerves in patients with diabetic peripheral neuropathy and controls.Protein profiling revealed 265 differentially expressed proteins in the diabetic peripheral neuropathy group.Gene Ontology indicated that differentially expressed proteins were mainly enriched in myelination and mitochondrial oxidative phosphorylation.A real-time polymerase chain reaction assay performed to validate the circRNA sequencing results yielded 11 differentially expressed circRNAs.circ_0002538 was markedly downregulated in patients with diabetic peripheral neuropathy.Further in vitro experiments showed that overexpression of circ_0002538 promoted the migration of Schwann cells by upregulating plasmolipin(PLLP)expression.Moreover,overexpression of circ_0002538 in the sciatic nerve in a streptozotocin-induced mouse model of diabetic peripheral neuropathy alleviated demyelination and improved sciatic nerve function.The results of a mechanistic experiment showed that circ_0002538 promotes PLLP expression by sponging miR-138-5p,while a lack of circ_0002538 led to a PLLP deficiency that further suppressed Schwann cell migration.These findings suggest that the circ_0002538/miR-138-5p/PLLP axis can promote the migration of Schwann cells in diabetic peripheral neuropathy patients,improving myelin sheath structure and nerve function.Thus,this axis is a potential target for therapeutic treatment of diabetic peripheral neuropathy.

Cdk5 and aberrant cell cycle activation at the core of neurodegeneration

Neurodegenerative diseases are caused by the progressive loss of specific neurons.The exact mechanisms of action of these diseases are unknown,and many studies have focused on pathways related to abnormal accumulation and processing of proteins,mitochondrial dysfunction,and oxidative stress leading to apoptotic death.However,a growing body of evidence indicates that aberrant cell cycle re-entry plays a major role in the pathogenesis of neurodegeneration.The activation of the cell cycle in mature neurons could be promoted by several signaling mechanisms,including c-Jun N-terminal kinases,p38 mitogen-activated protein kinases,and mitogen-activated protein kinase/extracellular signal-regulated kinase cascades;post-translational modifications such as Tau-phosphorylation;and DNA damage response.In all these events,implicated Cdk5,a proline-directed serine/threonine protein kinase,seems to be responsible for several cellular processes in neurons including axon growth,neurotransmission,synaptic plasticity,neuronal migration,and maintenance of neuronal survival.However,under pathological conditions,Cdk5 dysregulation may lead to cell cycle re-entry in post-mitotic neurons.Thus,Cdk5 hyperactivation,by its physiologic activator p25,hyper-phosphorylates downstream substrates related to neurodegenerative diseases.This review summarizes factors such as oxidative stress,DNA damage response,signaling pathway disturbance,and Ubiquitin proteasome malfunction contributing to cell cycle re-entry in post-mitotic neurons.It also describes how all these factors are linked to a greater or lesser extent with Cdk5.Thus,it offers a global vision of the function of cell cycle-related proteins in mature neurons with a focus on Cdk5 and how this protein contributes to the development of Alzheimer’s disease,Parkinson’s disease,amyotrophic lateral sclerosis,and Huntington’s disease by cell cycle activation.

双膦酸盐修饰生长分化因子5促进MC3T3-E1细胞的成骨分化

背景:生长分化因子5作为骨形态发生蛋白的一员在软骨及骨组织修复领域表现出良好的应用潜力,增强生长分化因子5与骨组织的亲和力是提高蛋白使用效率的关键,因而开发具有骨靶向性的生长分化因子5蛋白具有重要意义。目的:利用双膦酸盐修饰生长分化因子5并探讨改性后蛋白对小鼠成骨前体细胞生长分化的影响。方法:采用化学交联法将生长分化因子5与帕米膦酸钠偶联,得到偶联帕米膦酸钠的生长分化因子5,采用傅里叶变换红外光谱、圆二色谱对其基团及结构进行表征,利用ELISA试剂盒测定生长分化因子5与磷酸钙的结合量以及生长分化因子5的体外释放量,用于表征其体外骨靶向性。将生长分化因子5(对照组)、偶联帕米膦酸钠的生长分化因子5(实验组)分别与成骨前体细胞MC3T3-E1共培养,以单独培养的细胞为空白对照,评价复合物对细胞增殖及分化等的影响。结果与结论:①红外光谱及圆二色谱结果表明,实验成功制备了双膦酸盐/生长分化因子5复合物且蛋白二级结构无显著变化;体外磷酸钙吸附结果表明,偶联帕米膦酸钠后,生长分化因子5与磷酸钙的吸附率增加了约1倍;在半胱氨酸存在条件下,偶联帕米膦酸钠的生长分化因子5的蛋白可释放出来;②CCK-8实验结果显示,实验组培养4,7 d的吸光度值高于对照组、空白对照组(P<0.0001);培养7 d后,实验组碱性磷酸酶表达明显高于对照组、空白对照组(P<0.0001);培养13 d后,实验组钙结节含量明显高于对照组、空白对照组(P<0.0001);qRT-PCR结果检测结果显示,培养7 d后,实验组碱性磷酸酶、骨钙素及Runx2的mRNA表达高于对照组、空白对照组(P<0.01,P<0.001,P<0.0001);③结果表明,双膦酸盐修饰有利于增强生长分化因子5与磷酸钙的结合能力,同时有利于增强其生物活性。

Early-Stage Cervical Cancerous Cell Detection from Cervix Images Using YOLOv5

Cervical Cancer(CC)is a rapidly growing disease among women throughout the world,especially in developed and developing countries.For this many women have died.Fortunately,it is curable if it can be diagnosed and detected at an early stage and taken proper treatment.But the high cost,awareness,highly equipped diagnosis environment,and availability of screening tests is a major barrier to participating in screening or clinical test diagnoses to detect CC at an early stage.To solve this issue,the study focuses on building a deep learning-based automated system to diagnose CC in the early stage using cervix cell images.The system is designed using the YOLOv5(You Only Look Once Version 5)model,which is a deep learning method.To build the model,cervical cancer pap-smear test image datasets were collected from an open-source repository and these were labeled and preprocessed.Then the YOLOv5 models were applied to the labeled dataset to train the model.Four versions of the YOLOv5 model were applied in this study to find the best fit model for building the automated system to diagnose CC at an early stage.All of the model’s variations performed admirably.The model can effectively detect cervical cancerous cell,according to the findings of the experiments.In the medical field,our study will be quite useful.It can be a good option for radiologists and help them make the best selections possible.

骨形态发生蛋白5对肝癌细胞生物学行为的影响及机制

目的 探究骨形态发生蛋白5(BMP5)对人肝癌细胞生物学行为的影响及机制。方法 将正常人肝细胞系L02与人肝癌细胞系Hep3B、Huh7置于37℃、5%CO_(2)细胞培养箱中培养,采用RT-qPCR法检测正常人肝细胞系L02与人肝癌细胞系Hep3B、Huh7中BMP5 mRNA表达。将肝癌细胞系Hep3B、Huh7分为对照(NC)组及低、中、高剂量组,低、中、高剂量组分别用1、10、100 ng/mL浓度的重组人骨形态发生蛋白5(rh-BMP5)处理细胞,通过CCK-8法检测细胞增殖,Annexin-V/PI流式细胞术检测细胞凋亡,划痕实验检测细胞迁移,Western blotting法检测增殖细胞核抗原PCNA和SMAD通路关键蛋白及其磷酸化(SMAD3、p-SMAD3)表达。结果 与正常肝细胞系L02相比,BMP5在肝癌细胞系Hep3B、Huh7中表达升高(P均<0.05)。与NC组相比,中、高剂量组rh-BMP5可促进Hep3B、Huh7细胞增殖和PCNA表达上调(P均<0.05),高剂量组100 ng/mL的rh-BMP5处理肝癌细胞24及48 h细胞凋亡率降低、迁移率增高(P均<0.05)。高剂量组与NC组相比,p-SMAD3表达升高(P<0.05)。结论 BMP5在肝癌细胞系中高表达,可能通过增强SMAD3活化促进肝癌细胞Hep3B、Huh7的增殖、迁移并抑制凋亡。