Fibrinogen-like protein 2/fibroleukin prothrombinase contributes to tumor hypercoagulability via IL-2 and IFN-γ

AIM： To examine the role of Fibrinogen-like protein 2 （fgl2）/fibroleukin in tumor development. Fgl2 has been reported to play a vital role in the pathogenesis in MHV-3 （mouse hepatitis virus） induced fulminant and severe hepatitis, spontaneous abortion, allo- and xenograft rejection by mediating ＂immune coagulation＂.METHODS： Tumor tissues from 133 patients with six types of distinct cancers and the animal tumor tissues from human hepatocellular carcinoma （HCC） model on nude mice （established from high metastasis HCC cell line MHCC97LM6） were obtained. RESULTS： HfgI2 was detected in tumor tissues from 127 out of 133 patients as well as tumor tissues collected from human HCC nude mice. Hfgl2 was highly expressed both in cancer cells and interstitial inflammatory cells including macrophages, NK cells, and CD8^＋ T lymphocytes and vascular endothelial cells. HfgI2 mRNA was localized in cells that expressed hfgI2 protein. Fibrin （nogen） colocalization with hfgl2 expression was determined by dual immunohistochemical staining. In vitro, IL-2 and IFN-γ, increased hfgl2 mRNA by 10-100 folds and protein expression in both THP-1 and HUVEC cell lines. One-stage clotting assays demonstrated that THP-1 and HUVEC cells expressing hfgl2 had increased procoagulant activity following cytokines stimulation. CONCLUSION： The hfgI2 contributes to the hypercoagulability in cancer and may induce tumor angiogenesis and metastasis via cytokine induction.

Fibrinogen-like protein 2 expression correlates with microthrombosis in rats with type 2 diabetic nephropathy

Fibrinogen-like protein 2 （fgl2）, a novel prothrombinase, is involved in microthrombosis. We examined fgl2 expression in the glomerular and tubulointerstitial capillaries and its correlation with microthromsis in rats with streptozocin-induced type 2 diabetic nephropathy. Our RT-PCR and immunoblotting analysis showed that fgl2 mRNA and protein levels were increased in microvascular endothelial cells of the glomeruli and renal interstitia at week 19 and became significantly elevated with the development of diabetic nephropathy （P 〈 0.01）. Fgl2 was not or only weakly expressed in the renal tissues of normal rats. Furthermore, a direct significant correlation （r = 0.543, P 〈 0.01） was found between fgl2 expression and microthrombotic capillaries in the renal tissues. Enzyme linked immunosorbent assays （ELISA） additionally showed that circulating TNF-α levels in rats with type 2 diabetes were significantly elevated and closely correlated with fgl2 expression （r = 0.871, P 〈 0.01）. Our results suggest that fgl2 may activate renal microthrombosis, thus contributing to glomerular hypertension and renal ischemia.

Fibrinogen-like protein 2 deficiency inhibits virus-induced fulminant hepatitis through abrogating inflammatory macrophage activation

BACKGROUND Heterogeneous macrophages play an important role in multiple liver diseases,including viral fulminant hepatitis(VFH).Fibrinogen-like protein 2(FGL2)is expressed on macrophages and regulates VFH pathogenesis;however,the underlying mechanism remains unclear.AIM To explore how FGL2 regulates macrophage function and subsequent liver injury during VFH.METHODS Murine hepatitis virus strain 3(MHV-3)was used to induce VFH in FGL2-deficient(Fgl2-/-)and wild-type(WT)mice.The dynamic constitution of hepatic macrophages was examined.Adoptive transfer of Fgl2-/-or WT bone marrowderived macrophages(BMDMs)into WT recipients with macrophages depleted prior to infection was carried out and the consequent degree of liver damage was compared.The signaling cascades that may be regulated by FGL2 were detected in macrophages.RESULTS Following MHV-3 infection,hepatic macrophages were largely replenished by proinflammatory monocyte-derived macrophages(MoMFs),which expressed high levels of FGL2.In Fgl2-/-mice,the number of infiltrating inflammatory MoMFs was reduced compared with that in WT mice after viral infection.Macrophage depletion ameliorated liver damage in WT mice and further alleviated liver damage in Fgl2-/-mice.Adoptive transfer of Fgl2-/-BMDMs into macrophage-removed recipients significantly reduced the degree of liver damage.Inhibition of monocyte infiltration also significantly ameliorated liver damage.Functionally,Fgl2 deletion impaired macrophage phagocytosis and the antigen presentation potential and attenuated the proinflammatory phenotype.At the molecular level,FGL2 deficiency impaired IRF3,IRF7,and p38 phosphorylation,along with NF-κB activation in BMDMs in response to viral infection.CONCLUSION Infiltrated MoMFs represent a major source of hepatic inflammation during VFH progression,and FGL2 expression on MoMFs maintains the proinflammatory phenotype via p38-dependent positive feedback,contributing to VFH pathogenesis.

Overexpression of fibrinogen-like protein 2 protects against T cell-induced colitis

AIMTo determine the effect of overexpression of fibrinogen-like protein 2 (FGL2) on regulatory T cell (Treg) and effector T (Teff) cell function on T cell-induced colitis in Rag1-/- mice.METHODSTreg and Teff cells from fgl2-/-, fgl2+/+, and fgl2Tg mice were purified by FACS. They were studied in vitro for immunosuppressive activity and cell proliferation and in vivo for their effects on the development and prevention of T cell-induced colitis in Rag1-/- mice.RESULTSIn vitro, fgl2Tg Treg had enhanced immunosuppressive activity, and fgl2Tg Teff had reduced proliferation to alloantigen stimulation. Transfer of Teff from C57Bl/6J mice (fgl2+/+) into Rag1-/- mice produced both clinical and histologic colitis with dense infiltrates of CD3+ T cells, crypt abscesses and loss of goblet cells. Fgl2Tg Treg prevented the development of T cell-induced colitis, whereas fgl2+/+ and fgl2-/- Treg were only partially protective. In mice that received fgl2Tg Treg, the ratio of Foxp3+ to CD3+ cells was increased both in the colon and in mesenteric lymph nodes, and Teff cell proliferation as determined by staining with Ki67 was reduced. Teff cells from fgl2Tg mice did not produce colitis.CONCLUSIONHere we show that fgl2Tg Teff are hypoproliferative and do not induce colitis. We further demonstrate that fgl2Tg Treg prevent colitis in contrast to fgl2+/+ Treg, which were only partially protective. These studies collectively provide a rationale for exploring the use of FGL2 or Treg expressing high levels of FGL2 in the treatment of inflammatory bowel disease.

Correlation of fibrinogen-like protein 2 with progression of acute pancreatitis in rats

AIM: To examine fibrinogen-like protein 2 (fgl2) expression during taurocholate-induced acute pancreatitis progression in rats and its correlation with pancreatic injury severity. METHODS: Forty-eight male Sprague-Dawley rats were randomly divided into the severe acute pancreatitis (SAP) group (n = 24) and the sham operation (SO) group (n = 24). Sodium taurocholate (4% at doses of 1 mL/kg body weight) was retrogradely injected into the biliopancreatic ducts of the rats to induce SAP. Pancreatic tissues were prepared immediately after sacrifice. At the time of sacrifice, blood was obtained for determination of serum amylase activity and isolation of peripheral blood mononuclear cells (PBMCs). Pancreatic tissue specimens were obtained for routine light microscopy including hematoxylin and eosin staining, and the severity of pancreatic injury was evaluated 1, 4 and 8 h after induction. Expression of fgl2 mRNA was measured in the pancreas and PBMCs using reverse transcription polymerase chain reaction. Expression of fgl2 protein was evaluated in pancreatic tissues using Western blotting and immunohistochemical staining. Masson staining was also performed to observe microthrombosis. RESULTS: At each time point, levels of fgl2 mRNAs in pancreatic tissues and PBMCs were higher (P < 0.05) in the SAP group than in the SO group. For pancreatic tissue in SAP vs SO, the levels were: after 1 h, 3.911 ± 1.277 vs 1.000 ± 0.673; after 4 h, 9.850 ± 3.095 vs 1.136 ± 0.609; and after 8 h, 12.870 ± 3.046 vs 1.177 ± 0.458. For PBMCs in SAP vs SO, the levels were: after 1 h, 2.678 ± 1.509 vs 1.000 ± 0.965; after 4 h, 6.922 ± 1.984 vs 1.051 ± 0.781; and after 8 h, 13.533 ± 6.575 vs 1.306 ± 1.179. Levels of fgl2 protein expression as determined by Western blotting and immunohistochemical staining were markedly up-regulated (P < 0.001) in the SAP group compared with those in the SO group. For Western blotting in SAP vs SO, the results were: after 1 h, 2.183 ± 0.115 vs 1.110 ± 0.158; after 4 h, 2.697 ± 0.090 vs 0.947 ± 0.361; and after 8 h, 3.258 ± 0.094 vs 1.208 ± 0.082. For immunohistochemical staining in SAP vs SO, the results were: after 1 h, 1.793 ± 0.463 vs 0.808 ± 0.252; after 4 h, 4.535 ± 0.550 vs 0.871 ± 0.318; and after 8 h, 6.071 ± 0.941 vs 1.020 ± 0.406. Moreover, we observed a positive correlation in the pancreas (r = 0.852, P < 0.001) and PBMCs (r = 0.735, P < 0.001) between fgl2 expression and the severity of pancreatic injury. Masson staining showed that microthrombosis (%) in rats with SAP was increased (P < 0.001) compared with that in the SO group and it was closely correlated with fgl2 expression in the pancreas (r = 0.842, P < 0.001). For Masson staining in SAP vs SO, the results were: after 1 h, 26.880 ± 9.031 vs 8.630 ± 3.739; after 4 h, 53.750 ± 19.039 vs 8.500 ± 4.472; and after 8 h, 80.250 ± 12.915 vs 10.630 ± 7.003.CONCLUSION: Microthrombosis due to fgl2 overexpression contributes to pancreatic impairment in rats with SAP, and fgl2 level may serve as a biomarker during early stages of disease.

Low-density lipoprotein receptor-related protein 2(LRP2)is required for lipid export in the midgut of the migratory locust,Locusta migratoria

Low-density lipoprotein receptor-related protein 2(LRP2)is a multifunctional endocytic receptor expressed in epithelial cells.In mammals,it acts as an endocytic receptor that mediates the cellular uptake of cholesterol-containing apolipoproteins to maintain lipid homeostasis.However,little is known about the role of LRP2 in lipid homeostasis in insects.In the present study,we investigated the function of LRP2 in the migratory locust Locusta migratoria(LmLRP2).The mRNA of LmLRP2 is widely distributed in various tissues,including integument,wing pads,foregut,midgut,hindgut,Malpighian tubules and fat body,and the amounts of LmLRP2 transcripts decreased gradually in the early stages and then increased in the late stages before ecdysis during the nymphal developmental stage.Fluorescence immunohistochemistry revealed that the LmLRP2 protein is mainly located in cellular membranes of the midgut and hindgut.Using RNAi to silence LmLRP2 caused molting defects in nymphs(more than 60%),and the neutral lipid was found to accumulate in the midgut and surface of the integument,but not in the fat body,of dsLmLRP2-treated nymphs.The results of a lipidomics analysis showed that the main components of lipids(diglyceride and triglyceride)were significantly increased in the midgut,but decreased in the fat body and hemolymph.Furthermore,the content of total triglyceride was significantly increased in the midgut,but markedly decreased in the fat body and hemolymph in dsLmLRP2-injected nymphs.Our results indicate that LmLRP2 is located in the cellular membranes of midgut cells,and is required for lipid export from the midgut to the hemolymphand fat body in locusts.

Exploration of cyclooxygenase-2 inhibitory peptides from walnut dreg proteins based on in silico and in vitro analysis

Walnut dreg protein hydrolysates(WDPHs)exhibit a variety of biological activities,however,the cyclooxygenase-2(COX-2)inhibitory peptide of WDPHs remain unclear.The aim of this study was to rapidly screen for such peptides in WDPHs through a combination of in silico and in vitro analysis.In total,1262 peptide sequences were observed by nano liquid chromatography/tandem mass spectrometry(nano LC-MS/MS)and 4 novel COX-2 inhibitory peptides(AGFP,FPGA,LFPD,and VGFP)were identified.Enzyme kinetic data indicated that AGFP,FPGA,and LFPD displayed mixed-type COX-2 inhibition,whereas VGFP was a non-competitive inhibitor.This is mainly because the peptides form hydrogen bonds and hydrophobic interactions with residues in the COX-2 active site.These results demonstrate that computer analysis combined with in vitro evaluation allows for rapid screening of COX-2 inhibitory peptides in walnut protein dregs.

GATA binding protein 2 mediated ankyrin repeat domain containing 26 high expression in myeloid-derived cell lines

BACKGROUND Thrombocytopenia 2,an autosomal dominant inherited disease characterized by moderate thrombocytopenia,predisposition to myeloid malignancies and normal platelet size and function,can be caused by 5’-untranslated region(UTR)point mutations in ankyrin repeat domain containing 26(ANKRD26).Runt related transcription factor 1(RUNX1)and friend leukemia integration 1(FLI1)have been identified as negative regulators of ANKRD26.However,the positive regulators of ANKRD26 are still unknown.AIM To prove the positive regulatory effect of GATA binding protein 2(GATA2)on ANKRD26 transcription.METHODS Human induced pluripotent stem cells derived from bone marrow(hiPSC-BM)INTRODUCTION Ankyrin repeat domain containing protein 26(ANKRD26)acts as a regulator of adipogenesis and is involved in the regulation of feeding behavior[1-3].The ANKRD26 gene is located on chromosome 10 and shares regions of homology with the primate-specific gene family POTE.According to the Human Protein Atlas database,the ANKRD26 protein is localized to the Golgi apparatus and vesicles,and its expression can be detected in nearly all human tissues[4].Moreover,UniProt annotation revealed that ANKRD26 is localized in the centrosome and contains coiled-coil domains formed by spectrin helices and ankyrin repeats[5,6].The most common disease related to ANKRD26 is thrombocytopenia 2(THC2),which is a rare autosomal dominant inherited disease characterized by lifelong mild-to-moderate thrombocytopenia and mild bleeding[7-9].Caused by the variants in the 5’-untranslated region(UTR)of ANKRD26,THC2 is defined by a decrease in the number of platelets in circulating blood and results in increased bleeding and decreased clotting ability[8,10].Due to the point mutations that occur in the 5’-UTR of ANKRD26,its negative transcription factors(TFs),Runt related transcription factor 1(RUNX1)and friend leukemia integration 1(FLI1),lose their repression effect[11].The persistent expression of ANKRD26 increases the activity of the mitogen activated protein kinase and extracellular signal regulated kinase 1/2 signaling pathways,which are potentially involved in the regulation of thrombopoietin-dependent signaling and further impair proplatelet formation by megakaryocytes(MKs)[11].However,the positive regulators of ANKRD26,which might be associated with THC2 pathology,are still unknown.

Neural Wiskott-Aldrich syndrome protein(N-WASP)promotes distant metastasis in pancreatic ductal adenocarcinoma via activation of LOXL2

Pancreatic ductal adenocarcinoma(PDAC)is one of the most aggressive solid malignancies.A specific mechanism of its metastasis has not been established.In this study,we investigated whether Neural Wiskott-Aldrich syndrome protein(N-WASP)plays a role in distant metastasis of PDAC.We found that N-WASP is markedly expressed in clinical patients with PDAC.Clinical analysis showed a notably more distant metastatic pattern in the N-WASP-high group compared to the N-WASP-low group.N-WASP was noted to be a novel mediator of epithelialmesenchymal transition(EMT)via gene expression profile studies.Knockdown of N-WASP in pancreatic cancer cells significantly inhibited cell invasion,migration,and EMT.We also observed positive association of lysyl oxidase-like 2(LOXL2)and focal adhesion kinase(FAK)with the N-WASP-mediated response,wherein EMT and invadopodia function were modulated.Both N-WASP and LOXL2 depletion significantly reduced the incidence of liver and lung metastatic lesions in orthotopic mouse models of pancreatic cancer.These results elucidate a novel role for N-WASP signaling associated with LOXL2 in EMT and invadopodia function,with respect to regulation of intercellular communication in tumor cells for promoting pancreatic cancer metastasis.These findings may aid in the development of therapeutic strategies against pancreatic cancer.

Polycytosine RNA-binding protein 1 regulates osteoblast function via a ferroptosis pathway in type 2 diabetic osteoporosis

BACKGROUND Recently,type 2 diabetic osteoporosis(T2DOP)has become a research hotspot for the complications of diabetes,but the specific mechanism of its occurrence and development remains unknown.Ferroptosis caused by iron overload is con-sidered an important cause of T2DOP.Polycytosine RNA-binding protein 1(PCBP1),an iron ion chaperone,is considered a protector of ferroptosis.AIM To investigate the existence of ferroptosis and specific role of PCBP1 in the development of type 2 diabetes.METHODS A cell counting kit-8 assay was used to detect changes in osteoblast viability under high glucose(HG)and/or ferroptosis inhibitors at different concentrations and times.Transmission electron microscopy was used to examine the morpho-logical changes in the mitochondria of osteoblasts under HG,and western blotting was used to detect the expression levels of PCBP1,ferritin,and the ferroptosis-related protein glutathione peroxidase 4(GPX4).A lentivirus silenced and overex-pressed PCBP1.Western blotting was used to detect the expression levels of the osteoblast functional proteins osteoprotegerin(OPG)and osteocalcin(OCN),whereas flow cytometry was used to detect changes in reactive oxygen species(ROS)levels in each group.RESULTS Under HG,the viability of osteoblasts was considerably decreased,the number of mitochondria undergoing atrophy was considerably increased,PCBP1 and ferritin expression levels were increased,and GPX4 expression was decreased.Western blotting results demonstrated that infection with lentivirus overexpressing PCBP1,increased the expression levels of ferritin,GPX4,OPG,and OCN,compared with the HG group.Flow cytometry results showed a reduction in ROS,and an opposite result was obtained after silencing PCBP1.CONCLUSION PCBP1 may protect osteoblasts and reduce the harm caused by ferroptosis by promoting ferritin expression under a HG environment.Moreover,PCBP1 may be a potential therapeutic target for T2DOP.

Glucokinase regulatory protein rs780094 polymorphism is associated with type 2 diabetes mellitus, dyslipidemia, non-alcoholic fatty liver disease, and nephropathy

In this editorial,we comment on the article by Liu et al published in the recent issue of the World Journal of Diabetes(Relationship between GCKR gene rs780094 polymorphism and type 2 diabetes with albuminuria).Type 2 diabetes mellitus(T2DM)is a chronic disorder characterized by dysregulated glucose homeostasis.The persistent elevated blood glucose level in T2DM significantly increases the risk of developing severe complications,including cardiovascular disease,re-tinopathy,neuropathy,and nephropathy.T2DM arises from a complex interplay between genetic,epigenetic,and environmental factors.Global genomic studies have identified numerous genetic variations associated with an increased risk of T2DM.Specifically,variations within the glucokinase regulatory protein(GCKR)gene have been linked to heightened susceptibility to T2DM and its associated complications.The clinical trial by Liu et al further elucidates the role of the GCKR rs780094 polymorphism in T2DM and nephropathy development.Their findings demonstrate that individuals carrying the CT or TT genotype at the GCKR rs780094 locus are at a higher risk of developing T2DM with albuminuria compared to those with the CC genotype.These findings highlight the importance of genetic testing and risk assessment in T2DM to develop effective preventive strategies and personalized treatment plans.

Targeting neuronal PAS domain protein 2 and KN motif/ankyrin repeat domains 1:Advances in type 2 diabetes therapy

This editorial summarizes the latest literature on the roles of neuronal PAS domain protein 2 and KN motif/ankyrin repeat domain 1 in type 2 diabetes(T2D).We highlight their involvement inβ-cell dysfunction,explore their potential as therapeutic targets,and discuss the implications for new treatment strategies.We offer valuable insights into relevant gene regulation and cellular mechanisms relevant for the targeted management of T2D.

Molecular Docking Studies of Botanical Beverage Mix Berries (LIFEGREENTM) against Breast Cancer Cells from Targeted Protein 1QQG, 7B5Q & 7B5O & Uterine Fibroid from Targeted Protein 2AYR, 6T41 & 3GRF

Fibroids, also called leiomyomas or myomas, are communal tumors of the muscle or uterine wall that affect about 20% of females who are of reproductive age. They can look as if singly or in clusters, and they often cease to grow after menopause. Fibroids can be classified as intramural, sub serosal, pedunculated, or submucosal based on where they are positioned in the uterus. Although fibroids are benign, they can grow quickly and cause a range of symptoms, such as pelvic pressure, heavy menstrual flow, and infertility. As a result, fibroids are a main reason behind hysterectomy surgeries. The majority of cases of breast cancer are ductal and lobular cancers, making it the second utmost common cancer in women international. Gene mutations like those in BRCA1 or BRCA2 knowingly raise the risk of breast and other cancers, typically with an earlier cancer onset. Cancer risk is influenced by a complex interplay of genetic abnormalities, environmental factors, and lifestyle selections. Further research into these relations is domineering. Although they are common in uterine leiomyomas, especially multiple leiomyomas, MED12 mutations do not significantly correlate with tumor size. These mutations have also been noticed in smooth muscle tumors and leiomyosarcomas, two other types of uterine cancer. The identification of MED12 mutations as the sole genetic abnormality originates in leiomyomas raises the opportunity of a role in the genesis of cancer. 10% - 15% of women who are of reproductive age have endometriosis, which grants serious difficulties because of its chronic nature and range of clinical symptoms. Even after effective surgeries, issues reoccur often, adding to the enormous financial burden. The effects of MED12 mutations have been experiential in recent studies examining the molecular causes of endometriosis-associated infertility, which have shown anomalies in cellular connections and signaling cascades. Computational techniques were used in this study to investigate LifeGreenTM’s potential to prevent uterine fibroids and breast cancer. The efficacy of LifeGreenTM as a preventive measure or a treatment for common gynecological matters was examined and modeled. We investigated the mechanisms underlying LifeGreenTM’s benefits in the treatment of uterine fibroids and breast cancer using computational techniques. Our research contributes to our understanding of its potential therapeutic benefits for women’s health.

FGL2在胃肠间质瘤的表达及临床意义

目的:探讨纤维蛋白原样蛋白2(FGL2)在胃肠道间质瘤(GIST)中的表达及临床意义。方法:运用免疫组化EnVision二步法检测158例GIST患者中FGL2蛋白的表达情况,分析其与临床病理参数之间的相关性,并探讨FGL2蛋白与GIST中肿瘤浸润性淋巴细胞(TILs)CD3^(+)T、CD8^(+)T、CD20^(+)T及CD68^(+)T细胞计数之间的关系。结果:FGL2蛋白在GIST中的表达与肿瘤大小、NIH危险度分级、坏死及核分裂象计数呈负相关(P<0.05),与患者性别及肿瘤发生部位差异无统计学意义(P>0.05)。在GIST中,FGL2表达与低CD3^(+)T细胞计数相关(P<0.05),而与CD8^(+)T、CD20^(+)T及CD68^(+)T细胞计数差异无统计学意义(P>0.05)。结论:FGL2蛋白参与GIST的发生和发展,其表达水平在一定程度上可预测GIST的恶性潜能。FGL2蛋白可能通过调节肿瘤微环境参与GIST的发生和发展。

下调HMGB2表达对肝癌LM3细胞上皮-间质转化的抑制作用及其AKT/mTOR信号通路机制

目的:探讨下调肝癌细胞中高迁移率族框蛋白2 (HMGB2)表达对肝癌细胞生物学行为及上皮-间质转化(EMT)进程的影响,并阐明其作用机制。方法:对数生长期的人肝癌LM3细胞分为阴性对照组和HMGB2 RNA干扰组(HMGB2 siRNA组),分别以Lipofectamin 2000为载体转染无关序列的RNA寡核苷酸(RNA oligo)和敲除HMGB2序列的RNA oligo。采用实时荧光定量PCR(RT-qPCR)法和Western blotting法检测2组细胞中HMGB2 mRNA和蛋白表达水平,分别采用细胞划痕实验和Transwell小室实验检测2组细胞的迁移和侵袭能力,采用Western blotting法检测2组细胞中E-钙黏蛋白(E-cadherin)、 N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白表达水平。结果:与阴性对照组比较,HMGB2 siRNA组细胞中HMGB2 mRNA和蛋白表达水平均明显降低(P<0.05),HMGB2 siRNA组细胞划痕愈合率明显降低(P<0.01),侵袭细胞数明显减少(P<0.01),细胞中E-cadherin蛋白表达水平明显升高(P<0.01),N-cadherin、Vimentin、mTOR、AKT和磷酸化AKT (p-AKT)蛋白表达水平明显降低(P<0.05或P<0.01)。结论:下调HMGB2的表达可降低肝癌LM3细胞迁移和侵袭能力并抑制EMT,其作用机制可能与参与调节AKT/mTOR通路相关蛋白表达有关。

多发性子宫肌瘤患者血清ANGPTL2、VASH1的表达及临床意义

目的 分析血清中血管生成素样蛋白2(ANGPTL2)和血管生成抑制蛋白1(VASH1)的表达水平,探讨其在多发性子宫肌瘤中的临床意义。方法 选取2018年1月至2022年1月于东南大学医学院附属南京同仁医院就诊的312例多发性子宫肌瘤患者为研究对象(观察组),同时期来本院体检的312名健康女性作为对照(对照组);酶联免疫吸附试验(ELISA)检测血清中ANGPTL2和VASH1水平;Pearson相关分析血清中ANGPTL2和VASH1水平的相关性;受试者工作特征(ROC)曲线分析血清中ANGPTL2和VASH1水平对多发性子宫肌瘤的诊断价值;Logistic回归分析多发性子宫肌瘤发生的影响因素。结果 观察组血清中ANGPTL2和VASH1水平显著高于对照组,差异有统计学意义(t=12.870、9.935,P<0.05);血清中ANGPTL2和VASH1水平与阴道不规则出血、肌瘤数量、最大肌瘤直径以及平均肌瘤体积有关,差异有统计学意义(P<0.05),而与年龄、月经情况、绝经情况、妊娠史、流产史、ER、PR以及肿瘤部位无关,差异无统计学意义(P>0.05);Pearson相关分析结果显示,多发性子宫肌瘤患者血清中ANGPTL2和VASH1水平呈正相关(r=5.440,P<0.05);ROC曲线分析结果显示,血清中ANGPTL2和VASH1水平联合诊断多发性子宫肌瘤的曲线下面积(AUC),效果较ANGPTL2和VASH1单一指标更好(P<0.05);Logistic回归分析结果显示,ANGPTL2、VASH1是多发性子宫肌瘤发生的影响因素(P<0.05)。结论 多发性子宫肌瘤患者血清中ANGPTL2和VASH1水平显著升高,两者联合可辅助诊断多发性子宫肌瘤。

脂肪间充质干细胞过表达骨形态发生蛋白2促进骨质疏松大鼠牙槽骨缺损修复

背景:颌骨在骨质疏松症中最容易受累,脂肪间充质干细胞和骨形态发生蛋白2具有促进骨质疏松症骨再生的效果,然而骨形态发生蛋白2修饰的脂肪间充质干细胞对骨质疏松症牙槽骨缺损的修复作用鲜有报道。目的:探究过表达骨形态发生蛋白2的脂肪间充质干细胞对骨质疏松大鼠牙槽骨缺损的修复作用。方法:①将过表达骨形态发生蛋白2基因的慢病毒感染大鼠脂肪间充质干细胞,通过检测绿色荧光蛋白和骨形态发生蛋白2表达进行鉴定;②切除卵巢建立骨质疏松大鼠模型,于上颌两侧第一磨牙位置制备3 mm×3 mm×3 mm的圆柱形缺损;③假手术组和骨质疏松组大鼠植入明胶海绵,脂肪间充质干细胞组植入空载体慢病毒感染的脂肪间充质干细胞与明胶海绵复合体,过表达骨形态发生蛋白2的脂肪间充质干细胞组植入过表达骨形态发生蛋白2的脂肪间充质干细胞与明胶海绵复合体,1个月后进行相关指标检测。结果与结论:①脂肪间充质干细胞组和过表达骨形态发生蛋白2的脂肪间充质干细胞组转染效率均达到70%以上;与脂肪间充质干细胞组相比,过表达骨形态发生蛋白2的脂肪间充质干细胞组骨形态发生蛋白2蛋白表达水平明显升高(P<0.05);②假手术组骨缺损区可见大量新骨生成;与假手术组相比,骨质疏松组有少量新骨生成,新骨面积明显减小,碱性磷酸酶、骨钙素及骨形态发生蛋白2 mRNA和蛋白水平明显降低;与骨质疏松组相比,脂肪间充质干细胞组和过表达骨形态发生蛋白2的脂肪间充质干细胞组有大量新骨生成,新骨面积明显增加,碱性磷酸酶、骨钙素及骨形态发生蛋白2 mRNA和蛋白水平明显升高,且过表达骨形态发生蛋白2的脂肪间充质干细胞组优于脂肪间充质干细胞组(均P<0.05);③结果表明,骨形态发生蛋白2在骨质疏松大鼠牙槽骨表达较少,过表达骨形态发生蛋白2的脂肪间充质干细胞能够促进骨质疏松大鼠牙槽骨缺损的成骨再生。

BMAL1减轻H_(2)O_(2)诱导的心肌细胞损伤机制研究

目的探讨脑和肌肉组织芳香烃受体核转运蛋白的类似蛋白1(BMAL1)通过核因子E2相关因子2(NRF2)调节活性氧(ROS)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体通路对过氧化氢(H_(2)O_(2))诱导的心肌细胞损伤的影响。方法体外培养H9c2细胞和BMAL1稳定过表达的H9c2细胞,建立H_(2)O_(2)诱导的H9c2细胞损伤模型,并将细胞分为对照(Control)组、H_(2)O_(2)组、BMAL1过表达(BMAL1-OE)组、BMAL1过表达+H_(2)O_(2)(BMAL1-OE+H_(2)O_(2))组、BMAL1过表达+NRF2抑制剂(BMAL1-OE+ML385)组、BMAL1过表达+NRF2抑制剂+H_(2)O_(2)(BMAL1-OE+ML385+H_(2)O_(2))组。采用CCK-8法检测细胞活力,荧光探针2’,7’-二氯荧光素二乙酸酯检测ROS生成,Western blot检测BMAL1、NRF2和NLRP3蛋白表达,酶联免疫吸附试验法检测白细胞介素(IL)-1β释放。结果与Control组相比,H_(2)O_(2)组H9c2心肌细胞活力减弱,ROS生成增多,BMAL1和NRF2蛋白表达水平降低,NLRP3蛋白表达水平升高,IL-1β释放增多(P<0.05);与H_(2)O_(2)组相比,BMAL1-OE+H_(2)O_(2)组H9c2心肌细胞活力升高,ROS生成减少,BMAL1和NRF2蛋白表达水平升高,NLRP3蛋白表达水平降低,IL-1β释放减少(P<0.05)。与BMAL1-OE+H_(2)O_(2)组相比,BMAL1-OE+ML385+H_(2)O_(2)组H9c2心肌细胞活力减弱,ROS生成增多,NLRP3蛋白表达水平升高,IL-1β释放增多(P<0.05)。结论BMAL1可减轻H_(2)O_(2)诱导的H9c2心肌细胞损伤,其机制可能与NRF2调节ROS/NLRP3炎症小体通路有关。

入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平与CHB肝纤维化严重程度的相关性及对疾病预后的预测价值

目的探讨入院时血清转化生长因子-β1(TGF-β1)、Smad同源蛋白2(Smad2)、Smad同源蛋白3(Smad3)及透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、层黏连蛋白(LN)、Ⅳ型胶原(CⅣ)水平与慢性乙型肝炎(CHB)肝纤维化严重程度的相关性及联合检测对疾病预后的预测价值。方法选取河南省中医院2021年3月至2022年3月收治的78例CHB肝纤维化患者作为研究组,选择同期78名健康体检者作为对照组。比较研究组和对照组及不同肝纤维化分期、不同炎症活动分级CHB肝纤维化患者入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平;分析入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平与肝纤维化分期、炎症活动分级的相关性。CHB肝纤维化患者治疗3个月后,根据患者预后分为预后良好和预后不良亚组,比较预后良好和预后不良患者入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平;分析入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平联合检测对CHB肝纤维化患者预后不良的预测价值。结果研究组入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ高于对照组(P<0.05);不同肝纤维化分期、炎症活动分级CHB肝纤维化患者入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ比较:S1<S2<S3<S4、G1<G2<G3<G4,差异有统计学意义(P<0.05);入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平与肝纤维化分期、炎症活动分级均呈正相关(P<0.05)。预后良好患者入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平均低于预后不良患者(P<0.05);入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平联合预测肝纤维化患者预后不良的曲线下面积(AUC)优于各指标单一检测(P<0.05)。结论CHB肝纤维化患者入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平均呈现高表达,且与肝纤维化分期、炎症活动分级密切相关,其联合检测对CHB肝纤维化患者预后有较高的预测价值,可用于评估CHB肝纤维化患者病情严重程度和预后,为制定针对性治疗措施提供参考。

2017年~2023年河南省猪圆环病毒2型的流行调查及遗传变异分析

为了解河南省猪圆环病毒2型(PCV2)的流行及变异情况,本研究采集该地区规模化猪场2017年1月~2023年6月939份表现为繁殖障碍和呼吸道症状的病猪血液或组织样品,采用PCR方法进行PCV2检测。结果显示,PCV2总阳性率为31.42%(295/939);2017年~2022年PCV2阳性率逐年降低,分别为58.65%(61/104)、49.48%(48/97)、28.57%(36/126)、16.15%(31/192)、11.52%(19/165)和7.87%(7/89),而2023年迅速上升,达到56.02%(93/166)。利用PCR扩增29份PCV2阳性样品的全基因组序列并测序,采用Meg Align分析PCV2流行株全基因组序列与Gen Bank中4株PCV2参考株全基因组序列的同源性;采用MEGA 11软件利用NJ法构建PCV2流行株ORF2基因与Gen Bank中24株PCV2参考株ORF2基因的系统发育树;采用Meg Align分析PCV2流行株Cap蛋白氨基酸序列的变异特征;采用RDP4和Sim Plot分析PCV2流行株全基因组的重组特征。全基因组同源性分析结果显示,本研究鉴定的PCV2全基因组序列之间的同源性为95.1%~100%,与PCV2参考株的同源性为93.7%~98.6%,其中与疫苗株PCV2a LG(HM038034)、PCV2b DBN-SX07-2(HM641752)和PCV2d SH(AY686763)的同源性分别为94.8%~96.2%、95.3%~98.6%和96.6%~97.8%,与来自丹麦的代表株PCV2c DK1980PMWSfree株(EU148503)的同源性为93.7%~95.1%。此外,两株2023年的PCV2流行株HN230522和HN231217与参考株的同源性仅为94.5%~97.9%。进化树结果显示,有19株PCV2流行株与PCV2d基因型参考株聚为一个分支,10株与PCV2b基因型参考株聚为一个分支。其中今年出现的PCV2流行株HN230522和HN231217株虽然属于PCV2d基因型,但处于一个相对独立的分支。Cap蛋白氨基酸序列分析结果显示,与疫苗株PCV2a LG(HM038034)、PCV2b DBN-SX07-2(HM641752)和PCV2d SH株(AY686763)相比,部分PCV2流行株在构象表位区(aa47~aa85和aa165~aa200)存在R^(48)H、A^(59)K/R、G^(85)D、P^(151)T、N^(178)S、R^(180)K和G^(197)S的突变,在基因型特异性结构域(aa190~aa191/aa206/aa210)存在K^(206)I的突变,且HN230522株在核定位信号区(NLS)(aa1~aa41)存在特有的V^(30)L突变,HN231217株在基因型特异性结构域(aa89~aa91)存在特有的^(89)RTV^(91)残基。重组分析结果显示,有高达65.52%(19/29)的PCV2流行株存在疑似的重组片段,且重组片段全部位于ORF1中。上述结果表明,今年以来河南省猪场PCV2阳性率快速上升,且流行株出现了较大变异,应加强对PCV2流行动态和遗传变异的监测。本研究为河南省PCV2分子流行病学、疫苗研究提供了参考依据。