Human epidermal growth factor receptor 2 expression level and combined positive score can evaluate efficacy of advanced gastric cancer

BACKGROUND Although treatment options for gastric cancer(GC)continue to advance,the overall prognosis for patients with GC remains poor.At present,the predictors of treatment efficacy remain controversial except for high microsatellite instability.AIM To develop methods to identify groups of patients with GC who would benefit the most from receiving the combination of a programmed cell death protein 1(PD-1)inhibitor and chemotherapy.METHODS We acquired data from 63 patients with human epidermal growth factor receptor 2(HER2)-negative GC with a histological diagnosis of GC at the Cancer Hospital,Chinese Academy of Medical Sciences between November 2020 and October 2022.All of the patients screened received a PD-1 inhibitor combined with chemotherapy as the first-line treatment.RESULTS As of July 1,2023,the objective response rate was 61.9%,and the disease control rate was 96.8%.The median progression-free survival(mPFS)for all patients was 6.3 months.The median overall survival was not achieved.Survival analysis showed that patients with a combined positive score(CPS)≥1 exhibited an extended trend in progression-free survival(PFS)when compared to patients with a CPS of 0 after receiving a PD-1 inhibitor combined with oxaliplatin and tegafur as the first-line treatment.PFS exhibited a trend for prolongation as the expression level of HER2 increased.Based on PFS,we divided patients into two groups:A treatment group with excellent efficacy and a treatment group with poor efficacy.The mPFS of the excellent efficacy group was 8 months,with a mPFS of 9.1 months after excluding a cohort of patients who received interrupted therapy due to surgery.The mPFS was 4.5 months in patients in the group with poor efficacy who did not receive surgery.Using good/poor efficacy as the endpoint of our study,univariate analysis revealed that both CPS score(P=0.004)and HER2 expression level(P=0.015)were both factors that exerted significant influence on the efficacy of treatment the combination of a PD-1 inhibitor and chemotherapy in patients with advanced GC(AGC).Finally,multivariate analysis confirmed that CPS score was a significant influencing factor.CONCLUSION CPS score and HER2 expression both impacted the efficacy of immunotherapy combined with chemotherapy in AGC patients who were non-positive for HER2.

Correlation between receptor-interacting protein 140 expression and directed differentiation of human embryonic stem cells into neural stem cells

Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.

Human umbilical cord mesenchymal stem cells attenuate diabetic nephropathy through the IGF1R-CHK2-p53 signalling axis in male rats with type 2 diabetes mellitus

diabetes mellitus(DM)is a disease syndrome characterized by chronic hyperglycaemia.A long-term high-glucose environment leads to reactive oxygen species(ROS)production and nuclear DNA damage.human umbilical cord mesenchymal stem cell(HUcMSC)infusion induces significant antidiabetic effects in type 2 diabetes mellitus(T2DM)rats.Insulin-like growth factor 1(IGF1)receptor(IGF1R)is important in promoting glucose metabolism in diabetes;however,the mechanism by which HUcMSC can treat diabetes through IGF1R and DNA damage repair remains unclear.In this study,a DM rat model was induced with high-fat diet feeding and streptozotocin(STZ)administration and rats were infused four times with HUcMSC.Blood glucose,interleukin-6(IL-6),IL-10,glomerular basement membrane,and renal function were examined.Proteins that interacted with IGF1R were determined through coimmunoprecipitation assays.The expression of IGF1R,phosphorylated checkpoint kinase 2(p-CHK2),and phosphorylated protein 53(p-p53)was examined using immunohistochemistry(IHC)and western blot analysis.Enzyme-linked immunosorbent assay(ELISA)was used to determine the serum levels of 8-hydroxydeoxyguanosine(8-OHdG).Flow cytometry experiments were used to detect the surface markers of HUcMSC.The identification of the morphology and phenotype of HUcMSC was performed by way of oil red“O”staining and Alizarin red staining.DM rats exhibited abnormal blood glucose and IL-6/10 levels and renal function changes in the glomerular basement membrane,increased the expression of IGF1 and IGF1R.IGF1R interacted with CHK2,and the expression of p-CHK2 was significantly decreased in IGF1R-knockdown cells.When cisplatin was used to induce DNA damage,the expression of p-CHK2 was higher than that in the IGF1R-knockdown group without cisplatin treatment.HUcMSC infusion ameliorated abnormalities and preserved kidney structure and function in DM rats.The expression of IGF1,IGF1R,p-CHK2,and p-p53,and the level of 8-OHdG in the DM group increased significantly compared with those in the control group,and decreased after HUcMSC treatment.Our results suggested that IGF1R could interact with CHK2 and mediate DNA damage.HUcMSC infusion protected against kidney injury in DM rats.The underlying mechanisms may include HUcMSC-mediated enhancement of diabetes treatment via the IGF1R-CHK2-p53 signalling pathway.

Human bocavirus 1 and 2 genotype-specific antibodies for rapid antigen testing in pediatric patients with acute respiratory infections

Background Previous serological studies of human bocavirus(HBoV)1 could not exclude cross-reactivity with the other three HBoVs,particularly HBoV2.Methods To search for genotype-specific antibodies against HBoV1 and HBoV2,the divergent regions(DRs)located on the major capsid protein VP3 were defined through viral amino acid alignment and structure prediction.DR-deduced peptides were used as antigens to harvest corresponding anti-DR rabbit sera.To determine their genotype specificities for HBoV1 and HBoV2,these sera samples were used as antibodies against the antigens VP3 of HBoV1 and HBoV2(expressed in Escherichia coli)in western blotting(WB),enzyme-linked immunosorbent assay(ELISA),and bio-layer interferometry(BLI)assays.Subsequently,the antibodies were evaluated with clinical specimens from pediatric patients with acute respiratory tract infection by indirect immunofluorescence assay(IFA).Results There were four DRs(DR1–4)located on VP3 with different secondary and tertiary structures between HBoV1 and HBoV2.Regarding the reactivity with VP3 of HBoV1 or HBoV2 in WB and ELISA,high intra-genotype cross-reactivity of anti-HBoV1 or HBoV2 DR1,DR3,and DR4,but not anti-DR2,was observed.Genotype-specific binding capacity of anti-DR2 sera was confirmed by BLI and IFA,in which only anti-HBoV1 DR2 antibody reacted with HBoV1-positive respiratory specimens.Conclusion Antibodies against DR2,located on VP3 of HBoV1 or HBoV2,were genotype specific for HBoV1 and HBoV2,respectively.

SOX7靶向ERK1/2/PD-L1通路抑制结直肠癌血管生成

目的探讨性别决定区Y框蛋白7(SOX7)对结直肠癌血管生成的影响及潜在作用机制。方法应用免疫荧光检测结直肠癌患者组织样本中SOX7表达水平,之后通过裸鼠、转染SOX7 mimic的人结直肠癌细胞系SW480细胞和人脐静脉内皮细胞(HUVEC)共培养进一步研究。用Western-blot验证SOX7与ERK1/2/PD-L1对结直肠癌细胞的相关蛋白表达的影响。用CCK8检测SOX7与ERK1/2/PD-L1对HUVEC增殖的影响。通过体外内皮细胞成管实验测定SOX7与ERK1/2/PD-L1对肿瘤血管生成的影响。结果SOX7在人结直肠癌组织中表达被抑制(P<0.01),同时SOX7的过表达抑制了小鼠体内肿瘤生长(P<0.01)。SW480细胞中SOX7的过表达抑制了ERK1/2、c-Jun的表达,并在ERK1/2的激动剂Senkyunolide I的作用下上调了SW480细胞的ERK1/2、c-Jun蛋白表达(P<0.01),逆转了SOX7对SW480细胞中ERK1/2、c-Jun蛋白表达的影响(P<0.01)。HUVEC中SOX7抑制了PD-L1、V-EGFR2、p-PI3K、HIF-1α的蛋白表达,Senkyunolide I上调了HUVEC的PD-L1、V-EGFR2、p-PI3K、HIF-1α的蛋白表达,并逆转了SOX7对HUVEC中上述相关蛋白表达的影响(P<0.01)。PD-1/PD-L1 Inhibitor 3抑制了PD-L1、V-EGFR2、p-PI3K、HIF-1α的蛋白表达,SOX7过表达在PD-1/PD-L1 Inhibitor 3的影响下并没有表现出抑制作用。CCK8实验结果显示SOX7过表达显著抑制了HUVEC的增殖能力,Senkyunolide I作用下的两组HUVEC增殖能力较SOX7 NC组与SOX7 mimic组明显上升,PD-1/PD-L1 Inhibitor 3作用下的两组HUVEC增殖能力较SOX7 NC组与SOX7 mimic组明显下降,以上均有明显统计学差异(P<0.01)。成管实验结果显示SOX7过表达抑制了HUVEC的血管生成,Senkyunolide I强烈加速了血管生成,而PD-1/PD-L1 Inhibitor 3血管生成则被显著抑制,以上均有明显统计学差异(P<0.01)。结论SOX7通过ERK1/2/PD-L1通路抑制结直肠肿瘤的增殖和血管生成,SOX7可能是晚期CRC患者临床治疗中潜在的抗血管生成靶点。

胃癌患者病灶组织CK7、HER2、MLH1、MSH2及MSH6的表达变化研究

目的:探究胃癌患者病灶组织角蛋白7(CK7)、人表皮生长因子受体-2(HER2)、错配修复蛋白MutL同源物1(MLH1)、错配修复蛋白MutS同源物2(MSH2)及错配修复蛋白MutS同源物6(MSH6)的表达变化情况。方法:选取2020年2月—2023年1月苏州市相城人民医院收治的120例胃癌患者为研究对象,比较病灶组织及癌旁组织的CK7、HER2、MLH1、MSH2及MSH6的表达情况,并比较不同性别、年龄、TNM分期、分化程度、淋巴结转移情况及病灶位置者的病灶组织CK7、HER2、MLH1、MSH2及MSH6表达情况。结果:胃癌患者病灶组织CK7、HER2阳性率及MLH1、MSH2、MSH6表达缺失率显著高于癌旁组织,差异有统计学意义(P<0.05)。不同性别、年龄及病灶位置病灶组织的CK7、HER2、MLH1、MSH2及MSH6表达比较,差异无统计学意义(P>0.05);Ⅲ期、Ⅳ期病灶组织CK7、HER2阳性率高于Ⅰ期、Ⅱ期,MLH1、MSH2、MSH6表达缺失率均显著低于Ⅰ期、Ⅱ期,差异有统计学意义(P<0.05)。分化程度较低者病灶组织的CK7、HER2阳性率显著高于分化程度较高者,MLH1、MSH2及MSH6表达缺失率显著低于于分化程度较高者,差异有统计学意义(P<0.05)。有淋巴结转移者CK7、HER2阳性率显著高于无淋巴结转移者,MLH1、MSH2及MSH6表达缺失率显著低于无淋巴结转移者,差异有统计学意义(P<0.05)。结论:胃癌患者病灶组织的CK7、HER2、MLH1、MSH2及MSH6的表达相对异常,且不同TNM分期、分化程度及淋巴结转移情况者的差异较大。

Maintaining cholesterol homeostasis: Sterol regulatory element-binding proteins

The molecular mechanism of how hepatocytes maintain cholesterol homeostasis has become much more transparent with the discovery of sterol regulatory element binding proteins (SREBPs) in recent years. These membrane proteins aremembers of the basic helix-loop-helix-leucine zipper (bHLHZip) family of transcription factors. They activate the expression of at least 30 genes involved in the synthesis of cholesterol and lipids. SREBPs are synthesized as precursor proteins in the endoplasmic reticulum (ER), where they form a complex with another protein, SREBP cleavage activating protein (SCAP). The SCAP molecule contains a sterol sensory domain. In the presence of high cellular sterol concentrations SCAP confines SREBP to the ER. With low cellular concentrations, SCAP escorts SREBP to activation in the Golgi. There, SREBP undergoes two proteolytic cleavage steps to release the mature, biologically active transcription factor, nuclear SREBP (nSREBP). nSREBP translocates to the nucleus and binds to sterol response elements (SRE) in the promoter/enhancer regions of target genes. Additional transcription factors are required to activate transcription of these genes. Three different SREBPs are known, SREBPs-1a, -1c and -2. SREBP-1a and -1c are isoforms produced from a single gene by alternate splicing. SREBP-2 is encoded by a different gene and does not display any isoforms. It appears that SREBPs alone, in the sequence described above, can exert complete control over cholesterol synthesis, whereas many additional factors (hormones, cytokines, etc.) are required for complete control of lipid metabolism. Medicinal manipulation of the SREBP/SCAP system is expected to prove highly beneficial in the management of cholesterol-related disease.

多巴丝肼、普拉克索联合治疗帕金森病的效果及对PARK2、CKMT1A、Netrin-1的影响

目的探究多巴丝肼、普拉克索联合治疗帕金森病(PD)的效果及对人帕金森病蛋白2(PARK2)、线粒体肌酸激酶1A(CKMT1A)及神经轴突导向因子1(Netrin-1)的影响。方法选择2021年7月至2023年6月于新乡医学院第一附属医院治疗的PD患者108例为研究对象,依据随机数字表法分为试验组和对照组,每组54例。对照组行多巴丝肼治疗,试验组行多巴丝肼、普拉克索联合治疗。观察两组治疗前后PD严重程度,认知功能水平,睡眠障碍情况,血清PARK2、CKMT1A、Netrin-1水平和不良反应。结果治疗后,试验组统一PD评定量表(UPDRS)各分项得分及总分、匹兹堡睡眠质量指数量表(PSQI)评分均低于对照组(P<0.05),简易智力状态检查量表(MMSE)评分高于对照组(P<0.05)。试验组血清PARK2、Netrin-1水平均高于对照组(P<0.05),血清CKMT1A水平低于对照组(P<0.05)。试验组总有效率大于对照组(P<0.05)。两组不良反应发生率差异无统计学意义(P>0.05)。结论多巴丝肼、普拉克索联合治疗PD可缓解患者症状,提高其认知功能及睡眠质量,改善血清PARK2、CKMT1A、Netrin-1水平。

杜仲续断汤辅助治疗创伤性胫骨平台骨折患者的疗效分析

【目的】观察杜仲续断汤(由杜仲、续断、秦艽、木瓜、川芎、乳香、炙川乌、苏木、甘草等中药组成)辅助治疗对创伤性胫骨平台骨折患者术后临床症状改善和对血清人骨形态发生蛋白2(BMP-2)、碱性磷酸酶(ALP)、胰岛素样生长因子1(IGF-1)水平的影响。【方法】将80例创伤性胫骨平台骨折患者随机分为对照组和治疗组,每组各40例。2组患者均给予钢板内固定手术治疗,在此基础上,治疗组给予杜仲续断汤治疗,疗程为30 d,并对2组患者进行约24周随访调查。观察2组患者的骨痂生长情况以及治疗前后美国特种外科医院膝关节功能评分(HSS)及血清BMP-2、ALP、IGF-1等骨形成相关指标的变化情况,比较2组患者的临床疗效和不良事件发生情况。【结果】(1)疗效方面,经术后24周后的随访,治疗组和对照组的临床优良率分别为95.00%(38/40)和77.50%(31/40),组间比较,治疗组的疗效明显优于对照组(P<0.05)。(2)骨痂生长方面,治疗后,治疗组的不同骨痂生长量(包括少量、中量、大量)生成时间以及骨折愈合时间均较对照组明显缩短(P<0.05)。(3)骨形成相关指标方面,治疗后,2组患者血清BMP-2、ALP、IGF-1水平均较治疗前明显升高(P<0.05),且治疗组的升高幅度均明显优于对照组(P<0.01)。(4)膝关节功能方面,治疗后,2组患者的关节疼痛程度、行走能力判定、活动度判定、肌力判定、稳定性判定、屈曲畸形程度等各项HSS评分及其总分均较治疗前明显升高(P<0.05),且治疗组的升高幅度均明显优于对照组(P<0.01)。(5)不良事件方面,治疗组的不良事件发生率为5.00%(2/40),对照组为12.50%(5/40),组间比较,差异无统计学意义(P>0.05)。【结论】杜仲续断汤辅助治疗创伤性胫骨平台骨折患者,能有效改善患者术后临床症状,显著升高血清BMP-2、ALP、IGF-1等骨形成相关指标水平,缩短骨痂生成时间,促进膝关节功能恢复。

血清HE4、TM4SF1、HSP90α预测卵巢癌患者术后复发价值

目的:探究血清人附睾分泌蛋白4(HE4)、四次跨膜蛋白1(TM4SF1)、热休克蛋白90α(HSP90α)联合对卵巢癌(OC)患者术后复发的预测价值。方法:选取2018年10月-2020年12月在本院手术治疗的OC患者122例临床资料(OC组),根据术后3年随访复发情况分为未复发组(n=85)和复发组(n=37);同期在本院治疗的卵巢肿瘤良性患者102例(良性组)、本院健康体检正常女性115例为对照组。对比3组血清HE4、TM4SF1、HSP90α水平;logistic回归分析OC患者术后复发影响因素;受试者工作特征(ROC)曲线分析血清HE4、TM4SF1、HSP90α对OC术后复发的预测价值。结果:OC组血清HE4(158.49±41.27 pmol/L)、TM4SF1(171.28±42.33 ng/ml)、HSP90α(79.22±15.38 ng/ml)水平均高于良性组(34.53±8.39 pmol/L、111.18±25.46 ng/ml、58.19±10.24 ng/ml)和对照组(33.31±8.12 pmol/L、102.37±22.39 ng/ml、56.22±9.27 ng/ml),且复发组血清HE4、TM4SF1、HSP90α水平均高于未复发组(均P<0.05),而良性组与对照组水平均无差异(P>0.05)。logistic回归分析显示,FIGO分期III期、分化程度低、淋巴结转移、血清HE4、TM4SF1、HSP90α水平升高均为OC患者术后复发的影响因素。ROC曲线分析显示,HE4、TM4SF1、HSP90α单独预测OC患者术后复发的曲线下面积(AUC)分别为0.874、0.828、0.872,敏感度分别为89.2%、89.2%、89.2%,特异度分别为66.8%、69.2%、63.3%,3项联合预测OC患者术后复发的AUC为0.958,敏感度86.5%、特异度78.3%联合预测AUC最佳(P<0.05)。结论:OC术后复发患者术前血清HE4、TM4SF1、HSP90α水平均异常升高,且升高是OC患者术后复发的危险因素,3指标联合检测对OC患者术后复发有一定预测价值。

The in vitro reconstitution of nucleosome and its binding patterns with HMG1/2 and HMG14/17 proteins

Using atomic force microscopy (AFM), the dynamic process of the in vitro nucleosome reconstitution followed by slow dilution from high salt to low salt was visualized. Data showed that the histone octamers were dissociated from DNA at 1M NaCl. When the salt concentration was slowly reduced to 650 mMand 300 mM, the core histones bound to the naked DNA gradually. Once the salt concentration was reduced to 50 mM the classic 'beads-on-a-string' structure was clearly visualized. Furthermore, using the technique of the in vitro reconstitution ofnucleosome,the mono- and di- nucleosomes were assembled in vitro with both HS2core (-10681 to -10970 bp) and NCR2 (-372to -194 bp) DNA sequences in the 5'flanking sequence of human b-globin gene. Data revealed that HMG 1/2 and HMG 14/17 proteins binding to both DNA sequences are changeable following the assembly and disassembly of nucleosomes. We suggest that the changeable binding patterns of HMG 14/17 and HMG1/2 proteins with these regulatory elements may be critical in the process of nucleosome assembly, recruitment of chromatin-modifying activities, and the regulation of human b-globin gene expression.

MicroRNA-219a-5p和AFAP1L2对胰腺癌细胞的作用研究

目的探讨microRNA-219a-5p(miR-219a-5p)和肌动蛋白丝相关蛋白1相似蛋白2(AFAP1L2)对胰腺癌细胞的作用。方法分别采用实时荧光定量聚合酶链反应(qRT-PCR)和Western blotting法检测胰腺癌细胞株(PANC-1、BXPC-3、SW1990、Capan-1)及正常胰腺导管上皮细胞(HPDE)miR-219a-5p、AFAP1L2的表达,采用siRNA及过表达质粒下调或上调AFAP1L2表达,CCK-8法观察胰腺癌细胞株增殖变化。采用mimics或inhibitor上调或下调miR-219a-5p表达,CCK-8法观察胰腺癌细胞株增殖变化。流式细胞术检测不同干预条件下细胞凋亡和细胞周期。生物信息学预测两者可能具有靶向调控关系。qRT-PCR和Western blotting法检测AFAP1L2 mRNA及蛋白表达,荧光素酶实验观察二者碱基配对关系,进一步验证miR-219a-5p对AFAP1L2具有靶向调控作用。结果与HPDE细胞比较,胰腺癌细胞株中miR-219a-5p表达较低(P<0.05)。AFAP1L2 mRNA及蛋白在胰腺癌细胞株中的表达高于在HPDE细胞中的表达(P<0.05)。Capan-1或SW1990细胞培养48 h后,与空白对照组(NC组)比较,pCMV-EGFP-AFAP1L2组光密度(OD)值升高(P<0.05),siAFAP1L2组OD值下降(P<0.05),AFAP1L2对胰腺癌细胞增殖具有正向调控作用。Capan-1及SW1990细胞培养48 h后,与miR-NC组比较,miR-219a-5p mimics组OD值下降(P<0.05),miR-219a-5p inhibitor组OD值升高(P<0.05),miR-219a-5p表达对胰腺癌细胞增殖具有负向调控作用;上调miR-219a-5p表达,可诱导细胞S期阻滞,导致细胞凋亡增加。miR-219a-5p负向调控AFAP1L2表达;荧光素酶实验显示,miR-219a-5p与AFAP1L2的3'-URT互补结合,miR-219a-5p与AFAP1L2存在靶向调控关系。结论miR-219a-5p通过靶向调控AFAP1L2表达负向介导胰腺癌细胞增殖及凋亡。

Effects of Site-directed Mutagenesis of L469 in Helix-5 of Human Papillomavirus 16 L1 on Pentamer Formation

Located at the carboxyl terminal of the human papillomavirus major capsid protein L1, helix-5（h5） is cru-cial to L1 folding and pentamer formation. Site-directed mutagenesis of the leucine residue on site 469 into lysine, alanine, serine and glycine was performed to explore the effect of the resultant mutations on L 1 pentamer formation. The soluble yields of the L1 pentamers of the L469A and L469K mutants were nearly two fold higher than that of the wild type. Molecular dynamics simulation was then performed to reveal the intrinsic mechanisms involved in the improvement of L 1 pentamer yield. Accordingly, the secondary structures of h5, β-G2, β-B1, β-C, β-D, and β-F were altered. The altered structures improved the hydrophobic interaction between h5 and fl-core ＂jelly＂ and the stability of h5. The hydrophobic surface area of residue 469 was reduced by 50% relative to that of the wild type. The C--O group of residue 469 and C--N group of L470 were both exposed to the solvent in the L469A mutant. These modifications may account for the increased solubility and stability and the promotion of pentamer formation induced by the point mutation. Therefore, the changes in the hydrophobic properties of h5 and the core structure determined the pentamer formation and solubility. This study may assist the development of a cost-effective platform for the production of prophylactic virus-like particle vaccines.

Clinical signification of high-mobility group box 1protein(HMGB1) expression in infiltrating ductalcarcinoma breast tissue

Objective: Exploring the clinical signification of high-mobility group box 1 protein(HMGB1) expression in infiltrating ductal carcinoma(IDC) breast tissue. Methods: The expression of HMGB1 protein in IDC breast tissue was detected by immunohistochemistry, and the relations among size of tumour, lymph node metastasis, clinical staging, estrogen receptor(ER), progesterone receptor(PR) and human epidermal growth factor receptor 2(HER-2) were also analyzed. Results: Fortysix cases out of 60 cases of IDC breast tissue showed positive or strong positive HMGB1 expression(76.67%), statistical significance was observed between HMGB1 expression with clinical staging(P < 0.01), lymph node metastasis(P < 0.01), breast cancer ER(P < 0.05) and HER-2(P < 0.05), however same conclusion can not be drawn between HMGB1 with either size of tumour or PR expression(P > 0.05) in IDC breast tissue. Spearman analysis showed negative correlation between HMGB1 expression and ER, and positive correlation between HMGB1 expression and clinical staging, lymph node metastasis together with HER-2. Conclusion: It's promising that HMGB1 expression in IDC tissue can be one of biological indicators of poor prognosis.

The characterization of transmembrane protein 59-like(TMEM59L)reveals its role in the regulating the level of the GDI protein family

The characterization and functions of transmembrane protein 59-like(TMEM59L),a type I transmembrane protein,are not clearly understood until now.Some TMEM59L and fluorescent fusion proteins constructs were transfected in cell lines and liposomes,and their localization was observed.The effects of protein constructs were studied by fluorescence microscopy and western blotting.This study reports a novel function of human TMEM59L(hTMEM59L)related to the expression and location of some proteins.In addition,we report two novel splice variants of human TMEM59L(hTMEM59L).The localization of mutants of this protein,lacking a middle region,and a Cterminal deletion,markedly differed from that of full-length hTMEM59L.Intracellular movement assessment in living cells showed the localization of TMEM59L to vesicular structures in the Golgi bodies,and the cell membrane was observed in living cells.Overexpression of TMEM59L markedly increased the level of amyloid precursor protein because of TMEM59L-mediated inhibition of cell membrane transport but did not affect the expression of betasecretase 2.TMEM59L overexpression also significantly increased the levels of Rab GDP dissociation inhibitor alpha and Rab GDP dissociation inhibitor beta proteins.These results suggest that TMEM59L is involved in the packaging of acidic vesicles and thereby functions in the trafficking and processing of intracellular proteins.

人重组骨形态发生蛋白7通过下调半胱氨酸蛋白酶-1/焦孔素D信号通路抑制高糖诱导的近曲小管上皮细胞的焦亡

目的 观察人重组骨形态发生蛋白-7(recombinant human bone morphogenetic protein-7,rhBMP-7)对高糖诱导的人肾皮质近曲小管上皮细胞(human kidney-2,HK-2)的焦亡和炎症反应的作用。方法 体外培养高糖(high glucose,HG)刺激的HK-2细胞模拟糖尿病肾病模型。分为正常浓度葡萄糖对照组(Control)、高浓度葡萄糖组(HG)、Control+rhBMP-7组、HG+rhBMP-7组。采用免疫荧光方法检测HK-2细胞中E-钙黏蛋白(E-cadherin)、α平滑肌肌动蛋白(α-smooth muscle actin, α-SAM)、焦孔素D(gasderminD, GSDMD)蛋白和焦孔素E(gasderminE, GSDME)蛋白;TUNEL染色检测细胞死亡情况;蛋白质印迹法检测各组HK-2细胞中焦亡相关蛋白半胱氨酸蛋白酶-1(caspase-1)、半胱氨酸蛋白酶-4(caspase-4)、半胱氨酸蛋白酶-5(caspase-5)、 N-GSDMD和GAPDH的表达;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测各组HK-2细胞培养上清液中炎症因子白细胞介素-18(interleukin-18,IL-18)和白细胞介素-1β(interleukin-1β,IL-1β)的表达。结果 与Control组相比,高糖组HK-2细胞Tunel染色的阳性率明显增加,差异具有统计学意义(P<0.000 1),α-SMA蛋白和α-SMA mRNA表达量也显著增加(P<0.000 1),E-cadherin蛋白和Ecadherin mRNA表达量显著下降(P=0.000 6,P<0.000 1);HG组添加rhBMP-7后α-SMA蛋白和m RNA水平显著下降(P=0.011 7, P=0.002), E-cadherin蛋白和m RNA水平明显升高(P=0.001 5,P<0.000 1)。与对照组比较,HG组HK-2细胞GSDME/GSDMD蛋白的荧光信号明显增强。进而,HG组添加rhBMP-7后GSDME/GSDMD蛋白荧光信号显著减弱。同样,HG组HK-2细胞中焦亡相关蛋白caspase-1、caspase-4、caspase-5以及GSDMD的表达量显著高于对照组,差异明显(均P<0.001)。HG组添加rhBMP-7后焦亡相关蛋白的表达明显下降(分别P<0.001,P=0.002,P<0.001)。与对照组比较,HG组HK-2细胞培养上清液中IL-1β和IL-18蛋白浓度显著增加(均P<0.001)。进而导致rhBMP-7干预后HG组细胞上清液中IL-10和IL-1β蛋白浓度明显减少(P<0.001)。结论 rhBMP-7具有抑制HK-2细胞焦亡的作用,其通过调控caspase-1/GSDMD信号通路减缓高糖诱导的HK-2细胞焦亡。

蛋白激酶CK2β、P16蛋白、人乳头瘤病毒壳蛋白联合检测诊断宫颈癌的临床价值分析

目的:探讨宫颈癌患者采用蛋白激酶CK2β、P16蛋白、人乳头瘤病毒壳蛋白(HPVL-1)联合检测的临床诊断价值。方法:选取2020年6月—2022年1月单县中心医院收治的116例宫颈癌患者作为观察组,选取同时期收治的116例宫颈瘤变患者作为对照组。患者均接受病理、蛋白激酶CK2β、P16蛋白、HPVL-1检查,将临床综合检查结果作为金标准,分析蛋白激酶CK2β、P16蛋白、HPVL-1联合检测诊断宫颈癌的价值。结果:观察组HPVL-1阳性率低于对照组,P16、蛋白激酶CK2β阳性率高于对照组,差异有统计学意义(P<0.05)。HPVL-1、P16、蛋白激酶CK2β联合检测的灵敏度高于单一检测,漏诊率低于单一检测,差异有统计学意义(P<0.05)。结论:蛋白激酶CK2β、P16蛋白、HPVL-1联合检测诊断宫颈癌,灵敏度高,漏诊率低。

血清CTRP1、FGF21和ANGPTL3水平在2型糖尿病大血管病变中的诊断价值

目的观察血清C1q肿瘤坏死因子相关蛋白1(CTRP1)、成纤维细胞生长因子21(FGF21)和人血管生成素样蛋白3(ANGPTL3)水平在2型糖尿病大血管病变中的诊断价值。方法选取2020年1月至2022年6月在该院诊治的165例2型糖尿病患者作为2型糖尿病组,根据患者是否合并大血管病变分为大血管病变组(57例)和单纯糖尿病组(108例),另选取同期该院75例健康体检者作为健康对照组。采用单因素和多因素分析2型糖尿病发生大血管病变的影响因素,以及血清CTRP1、FGF21和ANGPTL3水平在2型糖尿病发生大血管病变中的诊断价值。结果2型糖尿病组血清CTRP1、FGF21和ANGPTL3水平均明显高于健康对照组,差异均有统计学意义(P<0.05),并且随着糖尿病控制程度升高而降低。大血管病变组餐后2 h血糖(2 h PG)、甘油三酯、低密度脂蛋白胆固醇(LDL-C)、颈动脉内膜中层厚度(IMT)、CTRP1、FGF21和ANGPTL3水平均明显高于单纯糖尿病组,差异均有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,2 h PG、TG、颈动脉IMT、CTRP1、FGF21和ANGPTL3水平升高是发生大血管病变的独立危险因素(P<0.05)。血清CTRP1、FGF21和ANGPTL3水平在2型糖尿病发生大血管病变中具有较高的诊断效能,3项指标联合检测的灵敏度为68.4%,特异度为97.2%,受试者工作特征曲线下面积为0.897,明显高于CTRP1(Z=3.152,P=0.002)、FGF21(Z=3.755,P<0.001)和ANGPTL3(Z=4.410,P<0.001)单项检测。结论血清CTRP1、FGF21和ANGPTL3是2型糖尿病的重要监测指标,其水平升高是发生大血管病变的独立危险因素,3项指标联合检测有助于提高对2型糖尿病大血管病变的诊断效能。

KEAP1-NRF2/HO-1通路介导LED红光促高糖诱导下人牙周膜干细胞成骨分化及减轻氧化损伤

目的探讨Kelch样ECH相关蛋白1-核因子E2相关因子2/血红素加氧酶-1(Kelch-like ECH associated protein 1-nuclear factor erythroid 2-related factor 2/heme oxygenase-1,KEAP1-NRF2/HO-1)通路介导发光二极管(light-emitting diode,LED)红光对高糖诱导下人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)成骨分化和氧化损伤的影响,为LED红光在细胞抗氧化损伤中的应用提供依据。方法流式细胞术、碱性磷酸酶(alkaline phosphatase,ALP)染色和茜素红染色鉴定hPDLSCs;高糖预处理hPDLSCs 48 h,用1、3、5 J/cm^(2)LED红光照射细胞,CCK-8实验选择促细胞增殖率高的辐射曝光量进行后续实验。将hPDLSCs分为对照组、高糖组、高糖+光照组;ALP染色、ALP活性检测、茜素红染色和半定量分析检测成骨分化能力,qRT-PCR和Western blot检测细胞成骨相关基因ALP、Runt相关转录因子2(runt-related transcription factor 2,RUNX2)、成骨细胞特异性转录因子(osterix,OSX)基因和蛋白表达;qRT-PCR检测相关抗氧化酶基因超氧化物歧化酶2(superoxide dismutase 2,SOD2)、过氧化氢酶(catalase,CAT)表达;荧光显微镜观察和流式细胞术测定细胞内活性氧簇(reactive oxygen species,ROS)水平;ELISA检测细胞上清液中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)水平。以NRF2特异性抑制剂ML385抑制NRF2通路,ALP染色、ALP活性检测细胞早期成骨分化能力,q RT-PCR检测早期成骨分化标志物ALP、RUNX2、OSX基因表达,Western blot检测细胞KEAP1、NRF2、HO-1蛋白表达水平。结果选择促高糖诱导下hPDLSCs增殖率最高的5 J/cm^(2)辐射曝光量进行后续实验(P<0.05)。5 J/cm^(2)LED红光促进高糖诱导下hPDLSCs的成骨分化(P<0.05),上调ALP、RUNX2、OSX的基因与蛋白表达(P<0.05),上调SOD2、CAT基因表达(P<0.05),降低细胞ROS水平(P<0.05),减少细胞上清液中TNF-α、IL-1β水平(P<0.05)。ML385抑制NRF2通路,细胞ALP活性降低(P<0.05),ALP、RUNX2、OSX基因表达下降(P<0.05),KEAP1蛋白表达上升(P<0.05),NRF2、HO-1蛋白表达下降(P<0.05)。结论LED红光可能通过KEAP1-NRF2/HO-1通路促进高糖诱导下hPDLSCs增殖和成骨分化,减轻氧化损伤。

子宫肌瘤患者血清Ang-2、HE4和ICF-1表达预测宫腔镜术后复发效果

目的:探讨子宫肌瘤患者血清促血管生成素-2(Ang-2)、人附睾分泌蛋白4(HE4)和胰岛素样生长因子-1(ICF-1)表达对宫腔镜术后复发的预测价值。方法:回顾性分析2019年1月-2020年10月本院妇产科接受治疗的126例子宫肌瘤患者为肌瘤组,体检健康妇女100例为对照组,肌瘤组采用宫腔镜手术治疗,术后随访1年,依据临床影像学及病理检查确诊肌瘤复发。比较肌瘤组术前和对照组体检当天血清Ang-2、HE4和ICF-1水平,根据随访结果判定子宫肌瘤复发者和未复发者,比较两类患者血清Ang-2、HE4和ICF-1水平,采用受术者工作特征曲线(ROC)分析Ang-2、HE4和ICF-1表达对宫腔镜术后肌瘤复发的预测价值。结果:血清Ang-2、HE4和ICF-1水平肌瘤组均高于对照组,且肌瘤组术后复发组(14例)高于未复发组(112例)(均P<0.05);血清Ang-2、HE4和ICF-1水平预测术后肌瘤复发的敏感度分别为83.8%、71.6%、51.4%,特异度分别为78.8%、75.0%、73.1%,3项联合检测预测术后复发的曲线下面积0.906,敏感度85.1%、特异度86.5%。结论:子宫肌瘤患者血清Ang-2、HE4和ICF-1水平异常表达,联合检测血清Ang-2、HE4和ICF-1水平对预测子宫肌瘤术后复发有较高价值。