Resveratrol inhibits pancreatic cancer proliferation and metastasis by depleting senescent tumor-associated fibroblasts

BACKGROUND Pancreatic cancer,a formidable gastrointestinal neoplasm,is characterized by its insidious onset,rapid progression,and resistance to treatment,which often lead to a grim prognosis.While the complex pathogenesis of pancreatic cancer is well recognized,recent attention has focused on the oncogenic roles of senescent tumor-associated fibroblasts.However,their precise role in pancreatic cancer remains unknown.Resveratrol is a natural polyphenol known for its multifaceted biological actions,including antioxidative and neuroprotective properties,as well as its potential to inhibit tumor proliferation and migration.Our current investigation builds on prior research and reveals the remarkable ability of resveratrol to inhibit pancreatic cancer proliferation and metastasis.AIM To explore the potential of resveratrol in inhibiting pancreatic cancer by targeting senescent tumor-associated fibroblasts.METHODS Immunofluorescence staining of pancreatic cancer tissues revealed prominent coexpression ofα-SMA and p16.HP-1 expression was determined using immunohistochemistry.Cells were treated with the senescence-inducing factors known as 3CKs.Long-term growth assays confirmed that 3CKs significantly decreased the CAF growth rate.Western blotting was conducted to assess the expression levels of p16 and p21.Immunofluorescence was performed to assess LaminB1 expression.Quantitative real-time polymerase chain reaction was used to measure the levels of several senescence-associated secretory phenotype factors,including IL-4,IL-6,IL-8,IL-13,MMP-2,MMP-9,CXCL1,and CXCL12.A scratch assay was used to assess the migratory capacity of the cells,whereas Transwell assays were used to evaluate their invasive potential.RESULTS Specifically,we identified the presence of senescent tumor-associated fibroblasts within pancreatic cancer tissues,linking their abundance to cancer progression.Intriguingly,Resveratrol effectively eradicated these fibroblasts and hindered their senescence,which consequently impeded pancreatic cancer progression.CONCLUSION This groundbreaking discovery reinforces Resveratrol's stature as a potential antitumor agent and positions senescent tumor-associated fibroblasts as pivotal contenders in future therapeutic strategies against pancreatic cancer.

Impact of STAT-signaling pathway on cancer-associated fibroblasts in colorectal cancer and its role in immunosuppression

Colorectal cancer(CRC)remains one of the most commonly diagnosed and deadliest types of cancer worldwide.CRC displays a desmoplastic reaction(DR)that has been inversely associated with poor prognosis;less DR is associated with a better prognosis.This reaction generates excessive connective tissue,in which cancer-associated fibroblasts(CAFs)are critical cells that form a part of the tumor microenvironment.CAFs are directly involved in tumorigenesis through different mechanisms.However,their role in immunosuppression in CRC is not well understood,and the precise role of signal transducers and activators of transcription(STATs)in mediating CAF activity in CRC remains unclear.Among the myriad chemical and biological factors that affect CAFs,different cytokines mediate their function by activating STAT signaling pathways.Thus,the harmful effects of CAFs in favoring tumor growth and invasion may be modulated using STAT inhibitors.Here,we analyze the impact of different STATs on CAF activity and their immunoregulatory role.

The Combined Effect of Lumenato and Ceramide in the Protection of Collagen Damage Induced by Neutrophils in Normal Human Dermal Fibroblasts

Introduction: Collagen is the primary structural protein fibroblasts produce in the skin’s extracellular matrix. Infiltration of neutrophils into the epidermis and dermis by exposure to UV causes collagen damage and contributes to photoaging. Methods: To study the combined effect of Lumenato and ceramide in preventing collagen-1 damage induced by phagocytes, we used co-cultures of normal human dermal fibroblasts (fibroblasts) and activated human neutrophils. The present study aimed to determine the protective effect of the combination of Lumenato and ceramide on fibroblast collagen-1 damage induced by neutrophils. Results: Lumenato (in the range of 6.5 - 208 μg/ml) or ceramide (in the range of 0.1 - 50 μM) inhibited the production of superoxides and MPO by TNFα-stimulated neutrophils, as well as the production of NO by LPS-stimulated macrophages in a dose-dependent manner. The combinations of Lumenato and ceramide, in low concentrations, caused synergistic prevention of fibroblasts’ collagen-1 damage induced by TNFα-activated neutrophils, detected by fluorescence immunostaining and WB analysis. MPO activity in the supernatants of the co-cultures was also synergistically inhibited. Adding Lumenato or ceramide singly or in combinations in these low concentrations to the fibroblast cultures did not affect the expression of collagen-1. The combinations of Lumenato or ceramide in these concentrations also caused a synergistic inhibition of NO production by activated macrophages. Conclusions: The results suggest that combining low concentrations of Lumenato and ceramide results in synergistic protection against fibroblasts’ collagen-1 damage induced by neutrophils, thus indicating their possible potential for enhanced skin health.

Lysine demethylase 5B transcriptionally regulates TREM1 in human cardiac fibroblasts

Background:A differential gene,triggering receptor expressed on myeloid cells 1(TREM1),was identified in blood sequencing datasets from myocardial infarction patients and healthy controls.Myocardialfibrosis following myocardial infarction significantly contributes to cardiac dysfunction.Objectives:This study aimed to unveil the intrinsic regulatory mechanism of TREM1 in myocardialfibrosis.Methods:Mimicking pathology by angiotensin II(Ang II)treatment of human cardiacfibroblasts(HCFs),the impacts of TREM1 knockdown on its proliferation,migration,and secretion of the pro-fibrotic matrix were identified.Using the Human Transcription Factor Database(HumanTFDB)website,lysine-specific demethylase 5B(KDM5B)was found to bind to the TREM1 promoter,which was further validated through luciferase reporter and chromatin immunoprecipitation(ChIP).By promoting KDM5B overexpression,its effect on the regulation of TREM1 was examined.Results:TREM1 knockdown suppressed the proliferation,migration,and secretion of the pro-fibrotic matrix in HCFs upon Ang II treatment.KDM5B bound to the TREM1 promoter and upregulated its transcriptional expression.Furthermore,KDM5B overexpression reversed the regulation of the above cellular phenotypes by TREM1 knockdown.Conclusion:This study sheds light on the positive regulation of TREM1 by KDM5B,demonstrating their role in promoting myocardialfibrosis.Thisfinding provides a theoretical foundation for understanding disease pathology and potentially advancing the development of new targeted therapies.

CRABP2 regulates infiltration of cancer-associated fibroblasts and immune response in melanoma

Finding biomarkers for immunotherapy is an urgent issue in cancer treatment.Cellular retinoic acid-binding protein 2(CRABP2)is a controversial factor in the occurrence and development of human tumors.However,there is limited research on the relationship between CRABP2 and immunotherapy response.This study found that negative correlations of CRABP2 and immune checkpoint markers(PD-1,PD-L1,and CTLA-4)were observed in breast invasive carcinoma(BRCA),skin cutaneous melanoma(SKCM),stomach adenocarcinoma(STAD)and testicular germ cell tumors(TGCT).In particular,in SKCM patients who were treated with PD-1 inhibitors,high levels of CRABP2 predicted poor prognosis.Additionally,CRABP2 expression was elevated in cancer-associated fibroblasts(CAFs)at the single-cell level.The expression of CRABP2 was positively correlated with markers of CAFs,such as MFAP5,PDPN,ITGA11,PDGFRα/βand THY1 in SKCM.To validate the tumor-promoting effect of CRABP2 in vivo,SKCM xenograft mice models with CRABP2 overexpression have been constructed.These models showed an increase in tumor weight and volume.Enrichment analysis indicated that CRABP2 may be involved in immunerelated pathways of SKCM,such as extracellular matrix(ECM)receptor interaction and epithelial-mesenchymal transition(EMT).The study suggests that CRABP2 may regulate immunotherapy in SKCM patients by influencing infiltration of CAFs.In conclusion,this study provides new insights into the role of CRABP2 in immunotherapy response.The findings suggest that CRABP2 may be a promising biomarker for PD-1 inhibitors in SKCM patients.Further research is needed to confirm these findings and to explore the clinical implications of CRABP2 in immunotherapy.

Unraveling the role of cancer-associated fibroblasts in colorectal cancer

Within the intricate milieu of colorectal cancer(CRC)tissues,cancer-associated fibroblasts(CAFs)act as pivotal orchestrators,wielding considerable influence over tumor progression.This review endeavors to dissect the multifaceted functions of CAFs within the realm of CRC,thereby highlighting their indispensability in fostering CRC malignant microenvironment and indicating the development of CAFs-targeted therapeutic interventions.Through a comprehensive synthesis of current knowledge,this review delineates insights into CAFsmediated modulation of cancer cell proliferation,invasiveness,immune evasion,and neovascularization,elucidating the intricate web of interactions that sustain the pro-tumor metabolism and secretion of multiple factors.Additionally,recognizing the high level of heterogeneity within CAFs is crucial,as they encompass a range of subtypes,including myofibroblastic CAFs,inflammatory CAFs,antigen-presenting CAFs,and vessel-associated CAFs.Innovatively,the symbiotic relationship between CAFs and the intestinal microbiota is explored,shedding light on a novel dimension of CRC pathogenesis.Despite remarkable progress,the orchestrated dynamic functions of CAFs remain incompletely deciphered,underscoring the need for continued research endeavors for therapeutic advancements in CRC management.

Effects of Serum Concentration, Synchronization Time and Confluence on the Cell-Cycle Synchronization Efficiency of Goat Fibroblasts

This study aims to evaluate the effect of serum concentration, synchronization time, and confluence degree on the synchronisation efficiency of goat fibroblast cycle. The results indicated that there was no difference in the percentage of nucleated fibroblasts in the G0/G1 stage between serum concentrations of 0.3% and 0.4% (83.89% and 82.69%, respectively, P > 0.05) as well as between serum concentrations of 0.2% and 0.5% (76.95% and 75.46%, respectively, P > 0.05). The percentage of nucleated fibroblasts in the G0/G1 stage was highest at the concentration of 0.3% and lowest in the control group (83.89% vs. 62.67%, P 0.05). The beneficial effect of high confluence was confirmed by the large percentage of nucleated fibroblasts at the G0/G1 stage. The 60% confluency was significantly lower than the 80% and 100% confluency (73.44%, 86.63%, and 87.17%, respectively, P < 0.05). The results indicate that the goat fibroblast cycle synchronization is the most effective at the serum concentration of 0.3%, 72 hours of synchronization and 100% confluency.

Calcitriol Suppressed Isoproterenol-induced Proliferation of Cardiac Fibroblasts via Integrinβ3/FAK/Akt Pathway

Objective Cardiac fibroblasts(CFs)proliferation and extracellular matrix deposition are important features of cardiac fibrosis.Various studies have indicated that vitamin D displays an anti-fibrotic property in chronic heart diseases.This study explored the role of vitamin D in the growth of CFs via an integrin signaling pathway.Methods MTT and 5-ethynyl-2′-deoxyuridine assays were performed to determine cell viability.Western blotting was performed to detect the expression of proliferating cell nuclear antigen(PCNA)and integrin signaling pathway.The fibronectin was observed by ELISA.Immunohistochemical staining was employed to evaluate the expression of integrinβ3.Results The PCNA expression in the CFs was enhanced after isoproterenol(ISO)stimulation accompanied by an elevated expression of integrin beta-3(β3).The blockade of the integrinβ3 with a specific integrinβ3 antibody reduced the PCNA expression induced by the ISO.Decreasing the integrinβ3 by siRNA reduced the ISO-triggered phosphorylation of FAK and Akt.Both the FAK inhibitor and Akt inhibitor suppressed the PCNA expression induced by the ISO in the CFs.Calcitriol(CAL),an active form of vitamin D,attenuated the ISO-induced CFs proliferation by downregulating the integrinβ3 expression,and phosphorylation of FAK and Akt.Moreover,CAL reduced the increased levels of fibronectin and hydroxyproline in the CFs culture medium triggered by the ISO.The administration of calcitriol decreased the integrinβ3 expression in the ISO-induced myocardial injury model.Conclusion These findings revealed a novel role for CAL in suppressing the CFs growth by the downregulation of the integrinβ3/FAK/Akt pathway.

Computed tomography-based radiomics predicts the fibroblastrelated gene EZH2 expression level and survival of hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma(HCC)is the most common subtype of liver cancer.The primary treatment strategies for HCC currently include liver transplantation and surgical resection.However,these methods often yield unsatisfactory outcomes,leading to a poor prognosis for many patients.This underscores the urgent need to identify and evaluate novel therapeutic targets that can improve the prognosis and survival rate of HCC patients.AIM To construct a radiomics model that can accurately predict the EZH2 expression in HCC.METHODS Gene expression,clinical parameters,HCC-related radiomics,and fibroblastrelated genes were acquired from public databases.A gene model was developed,and its clinical efficacy was assessed statistically.Drug sensitivity analysis was conducted with identified hub genes.Radiomics features were extracted and machine learning algorithms were employed to generate a radiomics model related to the hub genes.A nomogram was used to illustrate the prognostic significance of the computed Radscore and the hub genes in the context of HCC patient outcomes.RESULTS EZH2 and NRAS were independent predictors for prognosis of HCC and were utilized to construct a predictive gene model.This model demonstrated robust performance in diagnosing HCC and predicted an unfavorable prognosis.A negative correlation was observed between EZH2 expression and drug sensitivity.Elevated EZH2 expression was linked to poorer prognosis,and its diagnostic value in HCC surpassed that of the risk model.A radiomics model,developed using a logistic algorithm,also showed superior efficiency in predicting EZH2 expression.The Radscore was higher in the group with high EZH2 expression.A nomogram was constructed to visually demonstrate the significant roles of the radiomics model and EZH2 expression in predicting the overall survival of HCC patients.CONCLUSION EZH2 plays significant roles in diagnosing HCC and therapeutic efficacy.A radiomics model,developed using a logistic algorithm,efficiently predicted EZH2 expression and exhibited strong correlation with HCC prognosis.

17β-estradiol inhibits TGF-β-induced collagen gel contraction mediated by human Tenon fibroblasts via Smads and MAPK signaling pathways

AIM:To investigate the impact of 17β-estradiol on the collagen gels contraction(CGC)and inflammation induced by transforming growth factor(TGF)-βin human Tenon fibroblasts(HTFs).METHODS:HTFs were three-dimensionally cultivated in type I collagen-generated gels with or without TGF-β(5 ng/mL),17β-estradiol(12.5 to 100μmol/L),or progesterone(12.5 to 100μmol/L).Then,the collagen gel diameter was determined to assess the contraction,and the development of stress fibers was analyzed using immunofluorescence staining.Immunoblot and gelatin zymography assays were used to analyze matrix metalloproteinases(MMPs)and tissue inhibitors of metalloproteinases(TIMPs)being released into culture supernatants.Enzyme-linked immunosorbent assay(ELISA)and reverse transcription-quantitative polymerase chain reaction(RT-PCR)were used to detect interleukin(IL)-6,monocyte chemoattractant proteins(MCP)-1,and vascular endothelial growth factor(VEGF)in HTFs at the translational and transcriptional levels.The phosphorylation levels of Sma-and Mad-related proteins(Smads),mitogen-activated protein kinases(MAPKs),and protein kinase B(AKT)were measured by immunoblotting.Statistical analysis was performed using either the Tukey-Kramer test or Student’s unpaired t-test to compare the various treatments.RESULTS:The CGC caused by TGF-βin HTFs was significantly inhibited by 17β-estradiol(25 to 100μmol/L),and a statistically significant difference was observed when comparing the normal control group with 17β-estradiol concentrations exceeding 25μmol/L(P<0.05).The suppressive impact of 17β-estradiol became evident 24h after administration and peaked at 72h(P<0.05),whereas progesterone had no impact.Moreover,17β-estradiol attenuated the formation of stress fibers,and the production of MMP-3 and MMP-1 in HTFs stimulated by TGF-β.The expression of MCP-1,IL-6,and VEGF mRNA and protein in HTFs were suppressed by 100μmol/L 17β-estradiol(P<0.01).Additionally,the phosphorylation of Smad2 Smad3,p38,and extracellular signal-regulated kinase(ERK)were downregulated(P<0.01).CONCLUSION:17β-estradiol significantly inhibits the CGC and inflammation caused by TGF-βin HTFs.This inhibition is likely related to the suppression of stress fibers,inhibition of MMPs,and attenuation of Smads and MAPK(ERK and p38)signaling.17β-estradiol may have potential clinical benefits in preventing scar development and inflammation in the conjunctiva.

Exosomes from umbilical cord mesenchymal stromal cells promote the collagen production of fibroblasts from pelvic organ prolapse

BACKGROUND Pelvic organ prolapse(POP)involves pelvic organ herniation into the vagina due to pelvic floor tissue laxity,and vaginal structure is an essential factor.In POP,the vaginal walls exhibit abnormal collagen distribution and decreased fibroblast levels and functions.The intricate etiology of POP and the prohibition of trans-vaginal meshes in pelvic reconstruction surgery present challenges in targeted therapy development.Human umbilical cord mesenchymal stromal cells(hucMSCs)present limitations,but their exosomes(hucMSC-Exo)are promising therapeutic tools for promoting fibroblast proliferation and extracellular matrix remodeling.suppressed inflammation in POP group fibroblasts,stimulated primary fibroblast growth,and elevated collagen I(Col1)production in vitro.High-throughput RNA-seq of fibroblasts treated with hucMSC-Exo and miRNA sequencing of hucMSC-Exo revealed that abundant exosomal miRNAs downregulated matrix metalloproteinase 11(MMP11)expression.CONCLUSION HucMSC-Exo normalized the growth and function of primary fibroblasts from patients with POP by promoting cell growth and Col1 expression in vitro.Abundant miRNAs in hucMSC-Exo targeted and downregulated MMP11 expression.HucMSC-Exo-based therapy may be ideal for safely and effectively treating POP.

Clinical Observation of Recombinant Bovine Basic Fibroblast Growth Factor Eye Gel Combined with Tobramycin Dexamethasone Eye Drops in the Treatment of Dry Eye Syndrome in Patients after Cataract Surgery

Objective:To evaluate the therapeutic effect of recombinant bovine basic fibroblast growth factor(rbFGF)eye gel combined with tobramycin-dexamethasone(TOB-Dex)eye drops on dry eye syndrome(DES)after cataract surgery.Methods:86 patients with DES after cataract surgery,admitted from November 2021 to November 2023,were randomly divided into groups.The observation group included 43 patients treated with rbFGF eye gel combined with TOB-Dex eye drops.The reference group included 43 patients treated with TOB-Dex eye drops alone.Multiple indicators,including total effective rate and clinical symptom scores,were compared between the two groups.Results:The total effective rate in the observation group was higher than in the reference group(P<0.05).Before treatment,there were no differences in clinical symptom scores,serum factors,or disease severity scores between the two groups(P>0.05).Three weeks after treatment,the observation group had lower clinical symptom scores,serum factors,and disease severity scores compared to the reference group(P<0.05).The adverse reaction rate in the observation group was lower than in the reference group(P<0.05).Conclusion:rbFGF eye gel combined with TOB-Dex eye drops can improve the clinical efficacy for patients with DES after cataract surgery,alleviate disease symptoms,reduce inflammatory responses,and have fewer adverse reactions.

Bone marrow mesenchymal stem cell-derived exosomal microRNAs target PI3K/Akt signaling pathway to promote the activation of fibroblasts

BACKGROUND Fibroblast plays a major role in tendon-bone healing.Exosomes derived from bone marrow mesenchymal stem cells(BMSCs)can activate fibroblasts and promote tendon-bone healing via the contained microRNAs(miRNAs).However,the underlying mechanism is not comprehensively understood.Herein,this study aimed to identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets,and to verify their effects as well as mechanisms on fibroblasts.AIM To identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets and verify their effects as well as mechanisms on fibroblasts.METHODS BMSC-derived exosomal miRNAs data(GSE71241,GSE153752,and GSE85341)were downloaded from the Gene Expression Omnibus(GEO)database.The candidate miRNAs were obtained by the intersection of three data sets.TargetScan was used to predict potential target genes for the candidate miRNAs.Functional and pathway analyses were conducted using the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases,respectively,by processing data with the Metascape.Highly interconnected genes in the protein-protein interaction(PPI)network were analyzed using Cytoscape software.Bromodeoxyuridine,wound healing assay,collagen contraction assay and the expression of COL I andα-smooth muscle actin positive were applied to investigate the cell proliferation,migration and collagen synthesis.Quantitative real-time reverse transcription polymerase chain reaction was applied to determine the cell fibroblastic,tenogenic,and chondrogenic potential.RESULTS Bioinformatics analyses found two BMSC-derived exosomal miRNAs,has-miR-144-3p and hasmiR-23b-3p,were overlapped in three GSE datasets.PPI network analysis and functional enrichment analyses in the GO and KEGG databases indicated that both miRNAs regulated the PI3K/Akt signaling pathway by targeting phosphatase and tensin homolog(PTEN).In vitro experiments confirmed that miR-144-3p and miR-23b-3p stimulated proliferation,migration and collagen synthesis of NIH3T3 fibroblasts.Interfering with PTEN affected the phosphorylation of Akt and thus activated fibroblasts.Inhibition of PTEN also promoted the fibroblastic,tenogenic,and chondrogenic potential of NIH3T3 fibroblasts.CONCLUSION BMSC-derived exosomes promote fibroblast activation possibly through the PTEN and PI3K/Akt signaling pathways,which may serve as potential targets to further promote tendon-bone healing.

不同肠内营养制剂用于2型糖尿病患者对血糖及血清UA、PA、ALB的影响

目的探究康全力、能全力与瑞能三种不同肠内营养制剂用于2型糖尿病(T2DM)患者对血糖及血清尿酸(UA)、前白蛋白(PA)、白蛋白(ALB)等的影响。方法选取2020年1月至2023年3月于涟水县人民医院重症医学科内分泌与代谢性疾病科治疗的99例T2DM患者为研究对象,按非随机临床同期对照研究及患者自愿原则分为康全力组、能全力组和瑞能组,每组各33例。三组患者分别给予康全力、能全力和瑞能三种不同肠内营养制剂治疗,均连续治疗2个月,比较糖代谢、营养指标、血脂、炎症因子水平和不良反应发生情况。结果治疗后三组糖代谢指标均显著降低,其中康全力组患者治疗后糖化血红蛋白(HbA1c)、空腹血糖(FPG)、餐后2 h血糖(2hPG)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)与能全力组和瑞能组相比均显著降低(P<0.05);治疗后三组血清UA明显下降,PA和ALB水平明显升高(P<0.05),但组间差异无统计学意义(P>0.05);治疗后三组HDL-C均显著升高,康全力组LDL-C、TC和TG均显著降低(P<0.05),能全力组、瑞能组LDL-C、TC和TG未见显著改变(P>0.05),其中康全力组治疗后HDL-C与另外两组相比显著高,LDL-C与另外两组相比显著低,TC和TG与瑞能组相比显著低(P<0.05);治疗后三组IL-6、CRP水平与治疗前相比均显著降低(P<0.05),治疗后康全力组、能全力组IL-6和CRP水平与瑞能组相比显著低(P<0.05);治疗后康全力组、能全力组和瑞能组的不良反应发生总概率为12.12%、21.21%和15.15%,差异无统计学意义(P>0.05)。结论康全力在调节糖脂代谢、缓解炎症反应方面效果较其他肠内营养制剂更突出。

半固体培养法制备非洲猪瘟病毒pA104R蛋白的单克隆抗体

为了快速、高效制备非洲猪瘟病毒(ASFV)单克隆抗体,本研究通过大肠杆菌系统表达并纯化了ASFV重组蛋白pA104R。以ASFV重组蛋白pA104R为抗原,分别比较了CpG ODN联合氢氧化铝佐剂和常规弗氏佐剂两种免疫策略,并重点比较半固体培养法和常规有限稀释法来制备ASFV pA104R单克隆抗体的效率。结果显示,本研究获得了相对分子质量为3.5×104的ASFV重组可溶性蛋白pA104R,通过其与CpG ODN联合氢氧化铝佐剂免疫小鼠,在第21 d即可达到融合要求,本试验方法(重组蛋白pA104R与CpG ODN联合氢氧化铝佐剂免疫)较重组蛋白pA104R与常规弗氏佐剂免疫节省14 d时间。通过半固体培养法筛选单克隆的试验周期比有限稀释法缩短28 d,并减少了亚克隆的工作量。半固体培养法获得5株阳性杂交瘤细胞,挑选效价较高的3株(9A4、9H6、11F5)进行鉴定,重链均为IgG,轻链均为KAPPA。纯化后的3株单克隆抗体针对pA104蛋白和全病毒蛋白质的效价分别达1∶160000~1∶320000和1∶200~1∶400。本研究优选了CpG ODN联合氢氧化铝佐剂结合半固体培养法筛选pA104R的单克隆抗体,为单克隆抗体制备提供了快速高效的新策略。

抗生素溶杆菌中PAS-LuxR家族转录因子正向调控吩嗪类抗菌物质myxin的生物合成

溶杆菌Lysobacter是一类具有生防潜力的革兰氏阴性细菌。氮氧化吩嗪myxin是从抗生素溶杆菌Lysobacter antibioticus OH13中分离鉴定到的一种对病原细菌、真菌、卵菌等均具有较强拮抗活性的抗菌物质。我们已对myxin的生物合成机制进行了解析,但其生物合成的调控因子还鲜有报道。本研究在myxin生物合成基因簇上游鉴定到PAS-LuxR家族调控因子编码基因LaPhzR,LaPhzR敲除突变体丧失了产生myxin等吩嗪物质的能力,也丧失了对水稻条斑病菌RS105的拮抗能力,且myxin合成基因与解毒基因LaPhzX均不能表达。这些结果表明LaPhzR正向调控myxin的生物合成,且对myxin的生物合成是必需的。当在野生型菌株中过表达LaPhzR,myxin产量可提高1.6倍,这对myxin的开发应用具有重要意义。

帕金森病患者血清NPASDP-4,MBP水平表达与认知功能障碍及严重程度的诊断价值研究

目的探讨帕金森病患者血清神经元PAS结构域蛋白4(neuronal Per-Arnt-Sim domain protein 4,NPASDP-4)、髓鞘碱性蛋白(myelin basic protein,MBP)水平表达与认知功能障碍(cognitive impairment,CI)及严重程度的诊断价值研究。方法选取中国人民解放军联勤保障部队第九〇九医院收治的138例帕金森病患者为帕金森病组,同期该院体检中心的健康体检者69例为健康对照组,并根据是否发生CI以及其严重程度进一步将帕金森病组患者分为认知功能正常组(n=55)、轻度CI组(n=51)和痴呆组(n=32)。收集受试者一般资料;ELISA法检测血清NPASDP-4和MBP水平;相关性分析采用Spearman等级相关或Pearson线性相关;诊断价值分析采用ROC曲线;影响因素分析采用多因素Logistic回归。结果与健康对照组比较,帕金森病组血清NPASDP-4(6.75±0.48ng/ml vs2.38±0.31ng/ml),MBP(8.34±0.65μg/L vs 3.54±0.42μg/L)水平升高,差异具有统计学意义(t=68.751,55.761,均P<0.05)。认知功能正常组、轻度CI组、痴呆组H-Y分期比较,差异有统计学意义(χ2=7.788,P<0.05)。UPDRS-Ⅲ评分与认知功能正常组(41.95±10.36分)比较,轻度CI组(47.92±11.63分)、痴呆组(50.78±13.69分)评分升高,差异具有统计学意义(H=6.672,均P<0.05)。认知功能正常组、轻度CI组、痴呆组病程(4.28±0.54,4.71±0.58和5.16±0.63年)及血清NPASDP-4(5.89±0.40,6.83±0.55和8.12±0.54ng/ml),MBP(6.65±0.56,8.94±0.69和10.27±0.70μg/L)水平依次显著升高(H=24.114,207.950,355.594,均P<0.05),MMSE评分(28.47±0.94,24.51±1.35和17.09±2.57分)、MoCA评分(27.45±1.03,20.18±1.92和11.75±2.53分)、GPCOG总分(13.47±0.69,10.25±1.04和8.97±0.82分)依次显著降低(H=515.005,775.933,327.584,均P<0.05),差异具有统计学意义。帕金森病患者血清NPASDP-4,MBP水平均与病程(r=0.316,0.358)、H-Y分期(r=0.345,0.384)、UPDRS-Ⅲ评分(r=0.371,0.396)呈显著正相关(P<0.05),与MMSE评分(r=-0.468,-0.517)、MoCA评分(r=-0.504,-0.569)、GPCOG总分(r=-0.527,-0.538)呈显著负相关(均P<0.05)。血清NPASDP-4,MBP水平及二者联合诊断帕金森病患者CI的曲线下面积(AUC)分别为0.850,0.930和0.960,诊断帕金森病患者CI严重程度的AUC分别为0.866,0.803和0.933。H-Y分期中期[OR(95%CI):4.725(1.742~12.814)],H-Y分期晚期[OR(95%CI):5.083(1.919~13.464)]、UPDRS-Ⅲ评分[OR(95%CI):3.257(1.464~7.246)]、NPASDP-4[OR(95%CI):5.324(1.516~18.701)]和MBP[OR(95%CI):5.769(2.459~13.533)]是帕金森病患者CI的影响因素(均P<0.05);NPASDP-4[OR(95%CI):4.768(2.382~9.543)],MBP[OR(95%CI):5.846(3.141~10.882)]是帕金森病患者CI严重程度的影响因素(均P<0.05)。结论帕金森病患者血清NPASDP-4和MBP呈高水平,且均与CI及其严重程度密切相关,可能具有一定的临床诊断价值。

桂陈宣化汤联合t-PA溶栓对缺血性脑卒中患者血液流变学指标及神经营养因子影响

目的 观察桂陈宣化汤联合组织型纤溶酶原激活剂(t-PA)溶栓对缺血性脑卒中患者血液流变学指标及神经营养因子影响。方法 选择94例缺血性脑卒中患者,采用随机数字表法分为研究组与对照组各47例。对照组给予t-PA溶栓治疗,研究组在此基础上给予桂陈宣化汤,两组均连续治疗14 d。分别于治疗前后采用超声诊断仪检测脑平均血流速度(MBF)、平均血流量(CBF)、动态阻抗(DR)水平;采用血细胞分析仪检测全血黏度、血浆黏度、红细胞比容;采用ELISA法检测神经元特异性烯醇化酶(NSE)、神经营养因子(NTF)、神经生长因子(NGF)水平;采用脑卒中量表(NIHSS)、改良Rankin量表(MRS)评估患者脑卒中症状,评价临床疗效。结果 研究组总有效率为95.74%,高于对照组的80.85%(P <0.05)。治疗后,研究组MBF、CBF水平高于对照组(P <0.05),DR水平低于对照组(P <0.05);全血黏度、血浆黏度、红细胞比容低于对照组(P <0.05);NTF、NGF水平高于对照组(P <0.05),NSE水平低于对照组(P <0.05);NIHSS、MRS水平低于对照组(P <0.05)。结论 桂陈宣化汤联合t-PA溶栓治疗缺血性脑卒中患者疗效显著,能改善脑血流动力学、血液流变学指标水平,提高神经营养因子水平并缓解神经炎症,改善卒中症状。

不同尺寸石墨烯增强PA66纤维的效果分析

为提高PA66的力学性能,以石墨烯(GN)为基础填料,十二烷基苯磺酸钠(SDBS)作为表面活性剂,将GN均匀分散在SDBS的水溶液中,利用超声波粉碎技术,分别获得大尺寸(LGN)、中尺寸(MGN)、小尺寸(SGN)的GN,然后用熔融共混法制备出不同尺寸以及不同添加量的GN改性PA66,并对其性能进行了表征。研究结果表明:对于添加不同尺寸的GN,质量分数0.1%的SGN加入时,改性纤维的断裂强度提高至6.34 cN/dtex,较纯PA66提高了12.8%。同时SGN改性PA66的相对结晶度提升最大,为40.2%,相较于纯PA66提高了19.6%;对于不同添加量的SGN,SGN质量分数为0.1%时,改性纤维的断裂强度达到最大,力学性能提升最大。SGN的加入有利于异相成核,结晶速度相对加快。SGN改性PA66的最大分解速率温度为421.7℃,较纯PA66提高了9℃左右,说明SGN与PA66基体间发生了界面相互作用,具有热失重的延缓效果,加入SGN后PA66不易发生热分解。认为:添加一定量SGN能够更好提升PA66的各项性能。

聚酰胺(PA6)液相增粘反应技术的研究与开发

聚酰胺(PA6)作为重要的工程塑料之一,由于具有拉伸强度高、弹性模量大、耐磨损、自润滑和耐高温等特性,因此在汽车工业、电子行业、机械设备、海洋工程等行业得到广泛应用。本文主要针对聚酰胺(PA6)通过水解开环及固相增粘技术工艺存在反应效率低,工艺流程长,能耗高、分子量分布不均匀等缺点,通过PA6液相增粘反应技术小试试验研究,研究不同温度、停留时间及真空度下等条件下的增粘效果,其主要流程是缩聚反应完成之后的熔融物料直接进入液相增粘反应器中,通过双轴同向差速叶片连续搅拌不断的进行表面更新,同时低聚物、溶剂等小分子在更新表面溢出并被外部负压系统带走,完成增粘反应。聚酰胺(PA6)液相增粘反应技能耗显著降低,产品质量稳定,产品综合竞争力显著提高。