Meiotic nuclear divisions 1 suppresses the proliferation and invasion of pancreatic cancer cells via regulating H2A.X variant histone

Introduction:Among all malignant tumors of the digestive system,pancreatic carcinoma exhibits the highest mortality rate.Currently,prevention and effective treatment are urgent issues that need to be addressed.Methods:The study focused on meiotic nuclear divisions 1(MND1),integrating data from the Gene Expression Profiling Interactive Analysis(GEPIA)database with prognostic survival analysis.Simultaneously,experiments at cellular level were employed to demonstrate the effect of MND1 on the proliferation and migration of PC.The small-molecule inhibitor of MND1 was used to suppress the migration of PC cells by knocking down MND1 using small interfering RNA(siRNA)in Patu-8988 and Panc1 cell lines.Results:The results of Cell Counting Kit-8 indicated that the suppression of MND1 resulted in a decrease in cell proliferation.Wound healing and Transwell assays revealed that MND1 knockdown reduced cell migration and invasion.Flow cytometry revealed that inhibiting MND1 hindered the cell cycle.Furthermore,MND1 could stimulate the proliferation,migration,and invasion of Patu-8988 and Panc1 cells by increasing the expression of MND1.Notably,MND1 had a positive effect on H2AFX expression in PC cells.Elevated MND1 expression suggests the low overall survival rate of individuals diagnosed with PC.Conclusion:These findings suggest that MND1 has the potential to be a gene with the ability to accurately diagnose and treat PC.

Serpin家族H成员1在非小细胞肺癌中的表达及其作用机制研究

目的:分析Serpin家族H成员1(SERPINH1)在非小细胞肺癌(NSCLC)中的表达及其作用机制。方法:选取2022年1月-2023年12月于句容市人民医院进行肿瘤切除手术的60例NSCLC患者的肿瘤组织和癌旁组织进行研究。从癌症基因组图谱(TCGA)数据集获得NSCLC组织和正常组织的RNA测序(RNAseq)数据和相应的临床信息,免疫组化评估NSCLC组织中SERPINH1表达,并进行体外细胞实验分析SERPINH1对人肺腺癌细胞系A549细胞增殖、迁移和侵袭的影响。结果:与正常组织相比,SERPINH1 mRNA在NSCLC组织中表达显著上调。SERPINH1高表达患者总生存期和疾病特异性生存期短于SERPINH1低表达患者。NSCLC组织SERPINH1蛋白表达水平高于癌旁组织(P<0.001)。SERPINH1过表达组A549细胞增殖、迁移和侵袭能力高于空白组(P<0.001),SERPINH1敲低组A549细胞体外增殖、迁移和侵袭能力低于空白组(P<0.001)。结论:SERPINH1在NSCLC中高表达,与肿瘤增殖、转移密切相关,可作为治疗NSCLC的潜在靶点。

SERPINH1 promoted the proliferation and metastasis of colorectal cancer by activating PI3K/Akt/mTOR signaling pathway

BACKGROUND Serpin peptidase inhibitor clade H member 1(SERPINH1)was initially recognized as an oncogene implicated in various human malignancies.Nevertheless,the clinical relevance and functional implications of SERPINH1 in colorectal cancer(CRC)remain largely elusive.AIM To investigate the effects of SERPINH1 on CRC cells and its specific mechanism.METHODS Quantitative real-time polymerase chain reaction,western blotting analysis,The Cancer Genome Atlas data mining and immunohistochemistry were employed to examine SERPINH1 expression in CRC cell lines and tissues.A series of in-vitro assays were performed to demonstrate the function of SERPINH1 and its possible mechanisms in CRC.RESULTS SERPINH1 demonstrated elevated expression levels in both CRC cells and tissues,manifested at both mRNA and protein tiers.Elevated SERPINH1 levels correlated closely with advanced T stage,lymph node involvement,and distant metastasis,exhibiting a significant association with poorer overall survival among CRC patients.Subsequent investigations unveiled that SERPINH1 overexpression notably bolstered CRC cell proliferation,invasion,and migration in vitro,while conversely,SERPINH1 knockdown elicited the opposite effects.Gene set enrichment analysis underscored a correlation between SERPINH1 upregulation and genes associated with cell cycle regulation.Our findings underscored the capacity of heightened SERPINH1 levels to expedite G1/S phase cell cycle progression via phosphatidylinositol 3-kinase/AKT/mechanistic target of rapamycin pathway activation,thereby facilitating CRC cell invasion and migration.CONCLUSION These findings imply a crucial involvement of SERPINH1 in the advancement and escalation of CRC,potentially positioning it as a novel candidate for prognostic assessment and therapeutic intervention in CRC management.

Lysine-specific demethylase 1 inhibitor rescues the osteogenic ability of mesenchymal stem cells under osteoporotic conditions by modulating H3K4 methylation

Bone tissue engineering may be hindered by underlying osteoporosis because of a decreased osteogenic ability of autologous seed cells and an unfavorably changed microenvironment in these patients. Epigenetic regulation plays an important role in the developmental origins of osteoporosis; however, few studies have investigated the potential of epigenetic therapy to improve or rescue the osteogenic ability of bone marrow mesenchymal stem cells(BMMSCs) under osteoporotic conditions. Here, we investigated pargyline, an inhibitor of lysine-specific demethylase 1(LSD1), which mainly catalyzes the demethylation of the di- and mono-methylation of H3K4. We demonstrated that 1.5 mmol·Lpargyline was the optimal concentration for the osteogenic differentiation of human BMMSCs. Pargyline rescued the osteogenic differentiation ability of mouse BMMSCs under osteoporotic conditions by enhancing the dimethylation level of H3K4 at the promoter regions of osteogenesis-related genes. Moreover, pargyline partially rescued or prevented the osteoporotic conditions in aged or ovariectomized mouse models, respectively. By introducing the concept of epigenetic therapy into the field of osteoporosis, this study demonstrated that LSD1 inhibitors could improve the clinical practice of MSC-based bone tissue engineering and proposes their novel use to treat osteoporosis.

Expression of the B7 - related molecule B7 - H1 by glioma cells: a potential mechanism of immune paralysis

Human glioblastoma is a highly lethal tumor that is known for its immune inhibitory capabilities.B7-homologue l(B7-H 1),a recently identified homologue of B7.1/2(CD80/86),has been described to exert costimulatoryand immune regulatory functions.We investigated the expression and the functional activity of B7-H 1 in humanglioma cells in vitro and in vivo.Although lacking B7.1/2(CD80/86),all 12 glioma cel1 1ines constitutivelyexpressed B7-H1 mRNA and protein.Exposure to IFN-gamma strongly enhanced B7-H 1 expression.Im-

The Murine (C3H/He) Epidermal la^+ Dendritic Cells (la^+ DECs), Thy-1^+ Dendritic Cells (Thy-1^+DECs) and Aging

Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.

长链非编码RNA H 19调控小鼠睾丸间质细胞功能的机制

目的探讨长链非编码RNA H 19对小鼠睾丸间质细胞(LCs)合成分泌睾酮功能的影响及其可能的机制。方法采用原代小鼠LCs和TM3细胞为研究对象,细胞分为二甲基亚砜(DMSO)组、2-硝基丙烷(2-NP)处理组、si control组、si H19组、si Creb1组、Mimic control组、miR-181c-5p mimic组、Inhibitor control组、miR-181c-5p inhibitor组、si H19+Inhibitor control组和si H19+miR-181c-5p inhibitor组共11组。干扰H 19表达后,ELISA法检测其睾酮分泌水平的变化,CCK8法检测LCs增殖变化;生物信息学分析预测各基因的结合位点。RT-qPCR法和Western-blot检测各组小鼠LCs转染后各相关基因及蛋白表达的变化。结果(1)与si control组比较,si H19组LCs培养液睾酮水平及细胞增殖水平均显著下降(72 h后,P<0.01)。(2)各相关基因之间存在结合位点。(3)与DMSO组比较,2-NP组LCs的p-STAT1表达增加、H 19表达明显升高,而miR-181c-5p表达明显降低(P<0.01)。(4)与si control组比较,si H19处理可显著下调LCs的H 19、Creb1、HSD3B1和HSD3B6 RNA表达水平并显著上调miR-181c-5p表达水平(P<0.01),si Creb1处理可显著下调Creb1、HSD3B1和HSD3B6 mRNA表达水平(P<0.01);si H19组和si Creb1组Creb1、3β-HSD蛋白表达均明显降低(P<0.01);与Mimic control组比较,miR-181c-5p mimic组H 19、Creb1、HSD3B1和HSD3B6 mRNA表达明显降低,miR-181c-5p表达水平明显升高且Creb1、3β-HSD蛋白表达显著降低(P<0.01);与Inhibitor control组比较,miR-181c-5p inhibitor组的各相关基因及蛋白表达呈现与miR-181c-5p mimic组作用相反的效应;与si H19+Inhibitor control组比较,si H19+miR-181c-5p inhibitor组H 19表达显著升高,miR-181c-5p表达水平明显降低,Creb1、HSD的mRNA和蛋白表达均明显升高(P<0.01)。结论H 19低表达可通过抑制细胞增殖、miR-181c-5p表达升高、Creb1蛋白表达降低和3β-HSD蛋白表达降低导致LCs合成分泌睾酮能力降低。STAT1/H 19/miR-181c-5p/Creb1/3β-HSD通路可能在LCs功能调控中发挥重要作用。

H型高血压合并急性脑梗死患者血清L_(p)-PLA2、SAA、PECAM-1水平与病情程度和疾病预后的关系

目的 分析血清脂蛋白磷脂酶A2(L_(p)-PLA2)、淀粉样蛋白A(SAA)和血小板内皮细胞黏附分子-1(PECAM-1)水平与急性脑梗死(ACI)病情程度和疾病预后的相关性。方法 选择2021年7月—2022年6月海安市人民医院急诊科收治的高血压合并ACI患者180例,根据血清同型半胱氨酸(Hcy)水平分为观察组124例(H型高血压)和对照组56例(单纯高血压)。观察组再根据颈部超声检查结果分为非易损斑块亚组45例和易损斑块亚组79例,根据发病72 h内美国国立卫生院卒中量表(NIHSS)评分较入院时的变化值分为非早期神经功能恶化亚组(非END亚组)53例和END亚组71例,根据发病后90 d的改良Rankin量表(mRS)评分分成预后良好亚组89例和预后不良亚组35例。比较各组血清L_(p)-PLA2、SAA、PECAM-1水平。Pearson分析血清L_(p)-PLA2、SAA、PECAM-1水平与Hcy水平、Alberta卒中项目早期CT(ASPECTS)评分、急性生理与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分的相关性;绘制受试者工作特征曲线(ROC)分析血清L_(p)-PLA2、SAA、PECAM-1评估H型高血压合并ACI的效能。结果 血清L_(p)-PLA2、SAA、PECAM-1水平比较,观察组高于对照组(t/P=6.481/0.005、8.154/<0.001、7.462/<0.001),易损斑块亚组高于非易损斑块亚组(t/P=7.628/<0.001、8.319/<0.001、8.615/<0.001),END亚组高于非END亚组(t/P=7.854/<0.001、8.627/<0.001、9.825/<0.001),预后不良亚组高于预后良好亚组(t/P=7.915/<0.001、8.968/<0.001、8.763/<0.001)。Pearson分析显示,血清L_(p)-PLA2、SAA、PECAM-1水平与Hcy、APACHEⅡ评分均呈正相关(Hcy:r/P=0.691/<0.001、0.725/<0.001、0.674/<0.001;APACHEⅡ评分:r/P=0.591/<0.001、0.640/<0.001、0.683/<0.001),与ASPECTS评分呈负相关(r/P=-0.598/<0.001、-0.703/<0.001、-0.629/<0.001)。血清L_(p)-PLA2、SAA、PECAM-1及三者联合检测评估H型高血压合并ACI的曲线下面积(AUC)分别为0.681、0.709、0.754、0.924,三者联合检测评估效能高于单项检测(Z=6.863、7.418、7.905,P均<0.001)。结论 血清L_(p)-PLA2、SAA、PECAM-1在H型高血压合并ACI中呈高表达,与患者的颈动脉斑块稳定性、疾病严重程度和预后密切相关,三者联合检测的临床价值更高。

Possible mechanisms associated with immune escape and apoptosis on anti-hepatocellular carcinoma effect of Mu Ji Fang granules

BACKGROUND Hepatocellular carcinoma(HCC)is one of the most common digestive system cancers with high mortality rates worldwide.The main ingredients in Mu Ji Fang Granules(MJF)are alkaloids,flavonoids,and polysaccharides.MJF has been used in the clinical treatment of hepatitis,cirrhosis and HCC for more than 30 years.Few previous studies have focused on the mechanism of MJF on tumor immunology in the treatment of HCC.AIM To explore the mechanism of action of MJF on tumor immunology in the treatment of HCC.METHODS The absorbable ingredients of MJF were identified using Molecule Network related to High Performance Liquid Chromatography-Electron Spray Ionization-Time of Flight-Mass Spectrometry,and hub potential anti-HCC targets were screened using network pharmacology and pathway enrichment analysis.Forty male mice were randomly divided into the Blank,Model,and MJF groups(1.8,5.4,and 10.8 g/kg/d)following 7 d of oral administration.Average body weight gain,spleen and thymus indices were calculated,tumor tissues were stained with hematoxylin and eosin,and Interferon gamma(IFN-γ),Tumor necrosis factorα(TNF-α),Interleukin-2,aspartate aminotransferase,alanine aminotransferase,alpha-fetoprotein(AFP),Fas,and FasL were measured by Enzyme-linked Immunosorbent Assay.Relevant mRNA expression of Bax and Bcl2 was evaluated by Real Time Quantitative PCR(RTqPCR)and protein expression of Transforming growth factorβ1(TGF-β1)and Mothers against decapentaplegic homolog(SMAD)4 was assessed by Western blotting.The HepG2 cell line was treated with 10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL of MJF,and another 3 groups were treated with TGF-β1 inhibitor(LY364947)and different doses of MJF.Relevant mRNA expression of TNF-α,IFN-γ,Bax and Bcl2 was evaluated by RT-qPCR and protein expression of TGF-β1,SMAD2,p-SMAD2,SMAD4,and SMAD7 was assessed by Western blotting.RESULTS It was shown that MJF improved body weight gain and tumor inhibition rate in H22 tumorbearing mice,protected immune organs and liver function,reduced the HCC indicator AFP,affected immunity and apoptosis,and up-regulated the TGF-β1/SMAD signaling pathway,by increasing the relative expression of TGF-β1,SMAD2,p-SMAD2 and SMAD4 and decreasing SMAD7,reducing immune factors TNF-αand IFN-γ,decreasing apoptosis cytokines Fas,FasL and Bcl2/Bax,and inhibiting the effect of LY364947 in HepG2 cells.CONCLUSION MJF inhibits HCC by activating the TGF-β1/SMAD signaling pathway,and affecting immune and apoptotic cytokines,which may be due to MJF adjusting immune escape and apoptosis.

Hydrogen peroxide-induced changes in intracellular pH of guard cells precede stomatal closure

Epidermal bioassay demonstrated that benzylamine, a membrane-permeable weak base, can mimick hydrogen peroxide (H2O2) to induce stomatal closure, and butyric acid, a membrane-permeable weak acid, can partly abolish the H2O2-induced stomatal closure. Confocal pH mapping with the probe 5-(and-6)- carboxy seminaphthorhodafluor- 1 - acetoxymethylester (SNARF-1-AM) revealed that H2O2 leads to rapid changes in cytoplasmic and vacuolar pH in guard cells of Viola faba L, i. e. alkalinization of cytoplasmic areas occur red in parallel with a decrease of the vacuolar pH, and that butyric acid pretreatment can abolish alkalinization of cytoplasmic areas and acidification of vacuolar areas of guard cells challenged with H2O2. These results imply that the alkalinization of cytoplasm via efflux of cytosol protons into the vacuole in guard cells challenged with H2O2 is important at an early stage in the signal cascade leading to stomatal closure.

Numerical simulation of a triple-junction thin-film solar cell based on μc-Si_(1-x)Ge_x :H

In this paper, a-Si：H/a-SiGe：H/μc-SiGe：H triple-junction solar cell structure is proposed. By the analyses of mi- croelectronic and photonic structures （AMPS-1D） and our TRJ-F/TRJ-M/TRJ-B tunneling-recombination junction （TRJ） model, the most preferably combined bandgap for this structure is found to be 1.85 eV/1.50 eV/1.0 eV. Using more realistic material properties, optimized thickness combination is investigated. Along this direction, a-Si：H/a-SiGe：H/μc-SiGe：H triple cell with an initial efficiency of 12.09% （Voc = 2.03 V, FF = 0.69, Jsc = 8.63 mA/cm^2, area = 1 cm^2） is achieved in our laboratory.

Protein and gene expression characteristics of heterogeneous nuclear ribonucleoprotein H1 in esophageal squamous cell carcinoma

AIM To investigate the expression characteristics of heterogeneous nuclear ribonucleoprotein H1(HNRNPH1) m RNA and protein in cell lines and tissues of esophageal squamous cell carcinoma(ESCC). METHODS Western blotting was used to assess the expression of HNRNPH1 protein in seven ESCC cell lines and 30 paired fresh tissue specimens. The subcellular localization of HNRNPH1 was determined by immunofluorescence in ESCC cells. The RNA sequencing data from 87 patients with ESCC were obtained from the cancer genome atlas(TCGA), and the expression and clinical characteristics analysis of different transcript variants of HNRNPH1 were evaluated in this dataset. In addition, immunohistochemistry was carried out to detect the expression of HNRNPH1 protein in 125 patients.RESULTS The expression of HNRNPH1 protein varied across different ESCC cell lines. It was exclusively restricted to the nucleus of the ESCC cells. There are two transcript variants of the HNRNPH1 gene. Variant 1 was constitutively expressed, and its expression did not change during tumorigenesis. In contrast, levels of variant 2 were low in non-tumorous tissues and were dramatically increased in ESCC(P = 0.0026). The high levels of variant 2 were associated with poorer differentiated tumors(P = 0.0287). Furthermore, in paired fresh tissue specimens, HNRNPH1 protein was overexpressed in 73.3%(22/30) of neoplastic tissues. HNRNPH1 was significantly upregulated in ESCC, with strong staining in 43.2%(54/125) of tumor tissues and 22.4%(28/125) of matched non-cancerous tissues(P = 0.0005). Positive HNRNPH1 expression was significantly associated with poor tumor differentiation degree(P = 0.0337).CONCLUSION The different alternative transcript variants of HNRNPH1 exhibited different expression changes during tumorigenesis. Its m RNA and protein were overexpressed in ESCC and associated with poorer differentiation of tumor cells. These findings highlight the potential of HNRNPH1 in the therapy and diagnosis of ESCC.

Effect of miR-27b-3p and Nrf2 in human retinal pigment epithelial cell induced by high-glucose

AIM:To determine whether the microRNA-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRPE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically targeted.METHODS:hRPE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium kit.miR-27b-3p,Nrf2,NAD(P)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qPCR)and immunocytofluorescence(ICF),respectively.Western blot analyses were performed to determine nuclear and total Nrf2 protein levels.Nrf2,NQO1,and HO-1 expression levels by RT-qPCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral transfection.Finally,the expression level of those target genes was analyzed after treating hRPE cells with pyridoxamine.RESULTS:Persistent exposure to high glucose gradually suppressed hRPE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA levels.High glucose also promoted ROS release and inhibited cellular proliferation.Nrf2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p inhibitor.After treating hRPE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were suppressed.CONCLUSION:Nrf2 is a downstream target of miR-27b-3p.Furthermore,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.

神经纤毛因子1在骨折愈合中促进骨-血管偶联的作用

背景:神经纤毛因子1在骨折愈合过程中发挥重要作用,尤其是在促进骨-血管偶联方面。骨-血管偶联是骨折愈合过程中的关键事件之一,包括血管(尤其是H型血管)、新骨形成以及它们之间的关联。目的:回顾神经纤毛因子1在骨折愈合过程中促骨-血管偶联作用的研究进展。方法:在中国知网、PubMed数据库检索1982年1月至2024年4月发表的文献,以“Neuropilin-1,neuropilin 1,NRP1,neuropilin-1,NRP-1,Neuropilin 1,Nrp1,Nrp-1”为检索词,从中选择与骨折愈合、H型血管、动脉、骨细胞、骨-血管偶联关系紧密的文献43篇,增加手工检索的文献24篇,最终得到67篇文献。结果与结论:(1)在骨折愈合过程中骨-血管偶联的关键事件包括血管生成、新骨形成以及它们之间的关联,其中H型血管生成是骨折愈合过程中的关键事件之一,H型血管由血管内皮细胞以及周细胞组成;新骨形成主要是由成骨细胞、破骨细胞以及软骨细胞等骨细胞共同完成的;(2)神经纤毛因子1可以通过血管内皮生长因子和血小板衍生生长因子促进内皮细胞迁移、稳定,增强周细胞稳定性及其与内皮细胞之间的关联,从而促进血管再生;(3)神经纤毛因子1通过促进成骨细胞和软骨细胞形成、抑制破骨细胞生成来促进骨再生;(4)神经纤毛因子1可能是血管和骨偶联的关键因子之一;(5)局部应用神经纤毛因子1及其生物制剂,或将神经纤毛因子1、纳米载药系统以及高分子智能聚合材料等生物工程材料融合,为骨折治疗提供了新思路,有着广阔的应用前景。

Higher Viral Load and Prolonged Viral Shedding Period is Associated with Impaired Th17 Cell Response in Patients with H1N1 Influenza A

Objective To explore whether age,disease severity,cytokines and lymphocytes in H1N1 influenza A patients correlate with viral load and clearance.Methods Total of 70 mild and 16 severe patients infected with H1N1 influenza A virus were enrolled in this study.Results It was found that the patients under 14 years old and severe patients displayed significantly higher viral loads and prolonged viral shedding periods compared with the patients over 14 years old and mild patients,respectively(P < 0.05).Moreover,the patients under 14 years old and severe patients displayed significantly lower Th17 cell frequency than the patients over 14 years old and mild patients(P < 0.01).The viral shedding period inversely correlated with the frequency of IL-17+IFN-γ-CD4+ T cells.Additionally,the decreased concentration of serum TGF-β correlated with the decreased frequency of IL-17+IFN-γ-CD4+ T cells.Conclusions Both younger and severe patients are associated with higher viral loads and longer viral shedding periods,which may partially be attributed to the impaired Th17 cell response.

H1亚型猪流感病毒血凝素基因在昆虫细胞中的表达及其间接ELISA方法的初步建立

根据GenBank发表的H1亚型猪流感HA基因序列设计引物,扩增出HA基因片段,将其克隆到pFastBacGP67B杆状病毒载体上,筛选阳性重组转座载体pFastBacGP67B-H1,转化含有杆状病毒穿梭载体(bacmid)的DH10Bac感受态细胞,构建杆状病毒表达载体获得重组转座子(rBacmid-H1),在脂质体介导下转染sf9昆虫细胞,获得重组杆状病毒(rBV-H1),再感染细胞,收获目的蛋白。通过血凝试验、免疫印迹法、免疫组化分析表明该蛋白得到表达,且具有良好的生物学活性。利用表达的蛋白作为猪流感间接ELISA的抗原,初步建立H1亚型猪流感的间接ELISA检测方法,并对内蒙古、辽宁和黑龙江等地送检的93份猪血清进行了检测,阳性率为31.18%,为研制开发快速、准确、简便的H1亚型猪流感鉴别诊断试剂盒奠定基础。

nm23-H_1基因缺失人肺癌细胞株的筛选与鉴定

nm23-H1基因与肺癌的侵袭与转移密切相关,但是其作用的分子机制尚不清楚,为研究nm23-H1基因的功能,筛选并鉴定了nm23-H1基因缺失人肺癌细胞株及其生物学特性.应用Southernblot,RT-PCR和West-ernblot检测9株人肺癌细胞株中nm23-H1基因的存在状态及其生物学行为.结果发现发现人大肺癌细胞株L9981中存在nm23-H1等位基因的杂合性缺失,与其同源的NL9980及其它7株肺癌细胞株中nm23-H1基因均以杂合子的形式存在;并且L9981细胞株的增殖能力、克隆形成能力、体外侵袭力,裸鼠体内成瘤性及移植瘤肺转移的能力均显著高于NL9980.研究结果显示nm23-H1基因的缺失可能与L9981细胞株恶性表型和高转移潜能密切相关.

糖耐量正常人群OGTT 1h血糖与胰岛素敏感性和胰岛β细胞功能的关系

目的评价糖耐量正常(normal glucose tolerance,NGT)人群口服葡萄糖耐量试验(oral glucose tolerance test,OGTT)1 h血糖与胰岛素敏感性和胰岛β细胞功能的关系。方法收集101例NGT和76例糖耐量异常(impaired glucose tolerance,IGT)病人的临床资料;测定OGTT各时间点的血浆葡萄糖和胰岛素浓度;比较NGT 1 h血糖(1 hour postload plasma glucose,1-h PG)<8.6 mmol/L组、NGT 1-h PG≥8.6 mmol/L组和IGT组的胰岛素敏感性和胰岛β细胞功能,分析OGTT 1-h PG与胰岛素敏感性和胰岛β细胞功能的关系。结果 NGT 1-h PG≥8.6 mmol/L组的空腹血糖、1-h PG、1-h胰岛素及胰岛素敏感指数(homeostasis model assessment of insulin resistance,HOMA-IR)均显著高于NGT 1-h PG<8.6 mmol/L组;NGT 1-h PG≥8.6 mmol/L组的Matsuda指数、胰岛素分泌指数(ΔI3 0/ΔG3 0和ΔI6 0/ΔG6 0)均显著低于NGT 1-h PG<8.6 mmol/L组(P均<0.05)。但NGT 1-h PG≥8.6mmol/L组的空腹血糖、空腹胰岛素、1-h PG、1-h胰岛素、HOMA-IR、Matsuda指数、ΔI3 0/ΔG3 0和ΔI6 0/ΔG6 0与IGT组比较,差异无统计学意义(P均>0.05)。多元线性回归分析显示1-h PG与ΔI30/ΔG30、Matsuda指数呈线性负相关。结论在NGT人群中,OGTT 1-h PG≥8.6 mmol/L人群胰岛素敏感性和胰岛β细胞功能低于1-h PG<8.6 mmol/L组,1-h PG与胰岛素敏感性和胰岛β细胞功能相关。提示在NGT人群中,1-h PG浓度可作为评价胰岛素敏感性和胰岛β细胞功能的简易指标。

CD44V6和nm23-H_1蛋白在非小细胞肺癌中的表达及其临床意义

目的 研究 CD44 V6和 nm2 3- H1 蛋白在非小细胞肺癌 (NSCL C)中的表达水平及其临床意义。方法 采用免疫组化 SABC法检测 90例 NSCL C中 CD44 V6和 nm2 3- H1 蛋白表达。结果 CD44 V6和 nm2 3- H1 蛋白的阳性率分别为 5 1.1%、5 2 .2 % ;淋巴结转移组和无淋巴结转移组之间 CD44 V6和 nm2 3- H1 蛋白表达差异均有显著性 (P<0 .0 1) ;在 + 期与 + 期之间 CD44 V6蛋白表达差异有显著性 (P<0 .0 1) ;CD44 V6和 nm2 3- H1 蛋白在 NSCL C中的表达呈显著负相关 (P<0 .0 5 )。NSCL C中 CD44 V6和 nm2 3- H1 蛋白表达与病理类型、原发癌分期、有无远处转移无关。结论 NSCL C中 CD44 V6和 nm2 3- H1 蛋白表达与淋巴转移有关 ,可能具有预测 NSCL C淋巴转移的价值 ;CD44 V6和 nm2 3- H1 蛋白表达在 NSCL

过表达NHE1通过上调钙蛋白酶活性降低RAW264.7细胞ABCA1蛋白表达

目的探讨过表达钠氢交换体1(NHE1)对RAW264.7细胞三磷酸腺苷结合盒转运体A1(ABCA1)蛋白表达的影响。方法采用Ad NHE1腺病毒感染RAW264.7细胞,Western blot法检测NHE1-EGFP融合蛋白的表达,激光共聚焦显微镜技术检测NHE1-EGFP融合蛋白细胞内定位,酸负载p H回复检测NHE1活性,Western blot法检测NHE1-EGFP融合蛋白对RAW264.7细胞ABCA1蛋白水平及钙蛋白酶(calpain)活性的影响,加入calpain抑制剂N-乙酰基-L-亮氨酰-L-亮氨酰-L-正亮氨酸(ALLN),Western blot法检测肝脏X受体激动剂TO-901317诱导ABCA1蛋白表达水平的影响。结果 Ad NHE1腺病毒感染RAW264.7细胞后,高表达NHE1-EGFP融合蛋白,定位在细胞质及细胞膜。NHE1-EGFP融合蛋白可降低ABCA1蛋白的表达水平并提高calpain活性,而calpain抑制剂ALLN能阻断ABCA1蛋白表达水平降低。结论 NHE1过表达通过上调calpain活性而降低ABCA1蛋白表达水平。