AAV2-PDE6B restores retinal structure and function in the retinal degeneration 10 mouse model of retinitis pigmentosa by promoting phototransduction and inhibiting apoptosis

Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa.

Identification of hub genes associated with Helicobacter pylori infection and type 2 diabetes mellitus:A pilot bioinformatics study

BACKGROUND Helicobacter pylori(H.pylori)infection is related to various extragastric diseases including type 2 diabetes mellitus(T2DM).However,the possible mechanisms connecting H.pylori infection and T2DM remain unknown.AIM To explore potential molecular connections between H.pylori infection and T2DM.METHODS We extracted gene expression arrays from three online datasets(GSE60427,GSE27411 and GSE115601).Differentially expressed genes(DEGs)commonly present in patients with H.pylori infection and T2DM were identified.Hub genes were validated using human gastric biopsy samples.Correlations between hub genes and immune cell infiltration,miRNAs,and transcription factors(TFs)were further analyzed.RESULTS A total of 67 DEGs were commonly presented in patients with H.pylori infection and T2DM.Five significantly upregulated hub genes,including TLR4,ITGAM,C5AR1,FCER1G,and FCGR2A,were finally identified,all of which are closely related to immune cell infiltration.The gene-miRNA analysis detected 13 miRNAs with at least two gene cross-links.TF-gene interaction networks showed that TLR4 was coregulated by 26 TFs,the largest number of TFs among the 5 hub genes.CONCLUSION We identified five hub genes that may have molecular connections between H.pylori infection and T2DM.This study provides new insights into the pathogenesis of H.pylori-induced onset of T2DM.

应用Minigene剪接变异体分析技术诊断PMM2基因非经典剪接位点新变异的致病性

目的:研究Minigene剪接变异体分析技术在诊断磷酸甘露糖变位酶2(PMM2)相关先天性糖基化障碍(PMM2-CDG)中的价值,探讨磷酸甘露糖变位酶2(PMM2)基因剪接位点新变异对其转录产物的影响。方法:通过对1例PMM2-CDG患儿进行高通量测序查找可能的遗传学病因,利用Minigene剪接变异体分析技术,研究PMM2基因新剪接位点变异的致病性。根据美国医学遗传学与基因组学学会(ACMG)指南,判断新变异的致病性。结果:遗传学分析发现患儿系PMM2基因母源性c.691G>A(p.Val231Met)变异和父源性c.447+5G>A变异复合杂合子。Minigene剪接变异体分析发现:变异c.447+5G>A导致PMM2基因转录产物形成r.348_447del转录本,为致病性PMM2基因变异。患儿的临床特征为皮肤巩膜黄染,血清总胆红素、非结合胆红素和总胆汁酸明显升高,白蛋白明显降低,甲胎蛋白、铁蛋白和促甲状腺素等升高,对症支持治疗效果欠佳。结论:Minigene剪接变异体分析可为PMM2-CDG确诊和家系遗传咨询提供新的分子标记物,扩展了PMM2基因变异谱,为该病的临床诊治提供新的参考依据。

Vanillylacetone attenuates cadmium chloride-induced hippocampal damage and memory loss through upregulation of nuclear factor erythroid 2-related factor 2 gene and protein expression

Memory loss and dementia are major public health concerns with a substantial economic burden.Oxidative stress has been shown to play a crucial role in the pathophysiology of hippocampal damage-induced memory impairment.To investigate whether the antioxidant and anti-inflammatory compound vanillyla cetone(zingerone) can protect against hippocampal damage and memory loss induced by cadmium chloride(CdCl_(2)) administration in rats,we explo red the potential involvement of the nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathway,which is known to modulate oxidative stress and inflammation.Sixty healt hy male Wistar rats were divided into five groups:vehicle-treated(control),vanillylacetone,CdCl_(2),vanillylacetone+ CdCl_(2),vanillylacetone+ CdCl_(2)+ brusatol(a selective pharmacological N rf2inhibitor) groups.Vanillylacetone effectively attenuated CdCl_(2)-induced damage in the dental gyrus of the hippocampus and improved the memory function assessed by the Morris Water Maze test.Additionally,vanillylacetone markedly decreased the hippocampal tissue levels of inflammatory biomarkers(interleukin-6,tumor necrosis factor-α,intracellular cell adhesive molecules) and apoptosis biomarkers(Bax and cleaved caspase-3).The control and CdCl_(2)-treated groups treated with va nillylacetone showed reduced generation of reactive oxygen species,decreased malondialdehyde levels,and increased superoxide dismutase and glutathione activities,along with significant elevation of nuclear Nrf2 mRNA and protein expression in hippocampal tissue.All the protective effects of vanillylacetone we re substantially blocked by the co-administration of brusatol(a selective N rf2 inhibitor).Va nillylacetone mitigated hippocampal damage and memory loss induced by CdCl_(2),at least in part, by activating the nuclear transcription factor Nrf2.Additionally,vanillylacetone exerted its potent antioxidant and antiinflammatory actions.

To Analyze the Sensitivity of RT-PCR Assays Employing S Gene Target Failure with Whole Genome Sequencing Data during Third Wave by SARS-CoV-2 Omicron Variant

Introduction: Omicron is a highly divergent variant of concern (VOCs) of a severe acute respiratory syndrome SARS-CoV-2. It carries a high number of mutations in its spike protein hence;it is more transmissible in the community by immune evasion mechanisms. Due to mutation within S gene, most Omicron variants have reported S gene target failure (SGTF) with some commercially available PCR kits. Such diagnostic features can be used as markers to screen Omicron. However, Whole Genome Sequencing (WGS) is the only gold standard approach to confirm novel microorganisms at genetically level as similar mutations can also be found in other variants that are circulating at low frequencies worldwide. This Retrospective study is aimed to assess RT-PCR sensitivity in the detection of S gene target failure in comparison with whole genome sequencing to detect variants of Omicron. Methods: We have analysed retrospective data of SARS-CoV-2 positive RT-PCR samples for S gene target failure (SGTF) with TaqPath COVID-19 RT-PCR Combo Kit (ThermoFisher) and combined with sequencing technologies to study the emerged pattern of SARS-CoV-2 variants during third wave at the tertiary care centre, Surat. Results: From the first day of December 2021 till the end of February 2022, a total of 321,803 diagnostic RT-PCR tests for SARS-CoV-2 were performed, of which 20,566 positive cases were reported at our tertiary care centre with an average cumulative positivity of 6.39% over a period of three months. In the month of December 21 samples characterized by the SGTF (70/129) were suggestive of being infected by the Omicron variant and identified as Omicron (B.1.1.529 lineage) when sequence. In the month of January, we analysed a subset of samples (n = 618) with SGTF (24%) and without SGTF (76%) with Ct values Conclusions: During the COVID-19 pandemic, it took almost more than 15 days to diagnose infection and identify pathogen by sequencing technology. In contrast to that molecular assay provided quick identification with the help of SGTF phenomenon within 5 hours of duration. This strategy helps scientists and health policymakers for the quick isolation and identification of clusters. That ultimately results in a decreased transmission of pathogen among the community.

Analysis of SMOC2 gene variants in familial and nonfamilial primary open angle glaucoma Pakistani patients

AIM:To find out the association of secreted protein acidic and rich in cysteine(SPARC)-related modular calcium binding 2(SMOC2)gene variants rs2255680 and rs13208776 with genotypic and phenotypic characteristics in both familial and non-familial primary open angle glaucoma(POAG)patients.METHODS:A total of 212 POAG patients,comprising 124 familial and 88 non-familial,were enrolled.For genotyping the SMOC2 variant rs2255680,amplification refractory mutation system(ARMS)-polymerase chain reaction(PCR)method and PCR-restriction fragment length polymorphism(PCR-RFLP)were utilized for analyzing rs13208776 variant.RESULTS:The mean age of familial POAG patients was 50.92±9.12y,with 78 males and 46 females.The mean age of non-familial POAG patients was 53.14±13.44y,with 52 males and 36 females.The SMOC2 gene variant rs13208776 showed the significant association with POAG between familial and non-familial groups.The homozygous G/G variant was frequent among non-familial(60.2%)whereas the heterozygous G/A variant was more frequent in familial POAG patients(46%).There were significant differences in G/A variant between familial and non-familial glaucoma patients,and the risk was decreased to 0.53-fold in non-familial glaucoma patients[odds ratio(OR):0.53;95%confidence interval(CI):0.29-0.94;P=0.033]in codominant model.The risk was further reduced to 0.49-fold(95%CI:0.28-0.86;P=0.012)in dominant model for non-familial patients.No significant association of SMOC2 gene variant rs2255680 between familial and non-familial glaucoma patients was found in our population.The haplotype analysis showed the decreased risk for TA[OR:0.48(95%CI:0.29-0.79);P=0.004]and an increased risk for TG[OR=2.28(95%CI:1.22-4.25);P=0.01]haplotypes.CONCLUSION:Current findings show significant association of SMOC2 gene variant rs13208776 with POAG between familial and non-familial Pakistani patients.

Regulatory potential of soil available carbon,nitrogen,and functional genes on N_(2)O emissions in two upland plantation systems

Dynamic nitrification and denitrification processes are affected by changes in soil redox conditions,and they play a vital role in regulating soil N_(2)O emissions in rice-based cultivation.It is imperative to understand the influences of different upland crop planting systems on soil N_(2)O emissions.In this study,we focused on two representative rotation systems in Central China:rapeseed–rice(RR)and wheat–rice(WR).We examined the biotic and abiotic processes underlying the impacts of these upland plantings on soil N_(2)O emissions.The results revealed that during the rapeseed-cultivated seasons in the RR rotation system,the average N_(2)O emissions were 1.24±0.20 and 0.81±0.11 kg N ha^(–1)for the first and second seasons,respectively.These values were comparable to the N_(2)O emissions observed during the first and second wheat-cultivated seasons in the WR rotation system(0.98±0.25 and 0.70±0.04 kg N ha^(–1),respectively).This suggests that upland cultivation has minimal impacts on soil N_(2)O emissions in the two rotation systems.Strong positive correlations were found between N_(2)O fluxes and soil ammonium(NH_(4)^(+)),nitrate(NO_(3)^(–)),microbial biomass nitrogen(MBN),and the ratio of soil dissolved organic carbon(DOC)to NO_(3)^(–)in both RR and WR rotation systems.Moreover,the presence of the AOA-amoA and nirK genes were positively associated with soil N_(2)O fluxes in the RR and WR systems,respectively.This implies that these genes may have different potential roles in facilitating microbial N_(2)O production in various upland plantation models.By using a structural equation model,we found that soil moisture,mineral N,MBN,and the AOA-amoA gene accounted for over 50%of the effects on N_(2)O emissions in the RR rotation system.In the WR rotation system,soil moisture,mineral N,MBN,and the AOA-amoA and nirK genes had a combined impact of over 70%on N_(2)O emissions.These findings demonstrate the interactive effects of functional genes and soil factors,including soil physical characteristics,available carbon and nitrogen,and their ratio,on soil N_(2)O emissions during upland cultivation seasons under rice-upland rotations.

Pathogenesis of chronic enteropathy associated with the SLCO2A1 gene:Hypotheses and conundrums

Chronic enteropathy associated with the SLCO2A1 gene(CEAS)is a complex gastroenterological condition characterized by multiple ulcers in the small intestine with chronic bleeding and protein loss.This review explores the potential mechanisms underlying the pathogenesis of CEAS,focusing on the role of SLCO2A1-encoded prostaglandin transporter OATP2A1 and its impact on prostaglandin E2(PGE2)levels.Studies have suggested that elevated PGE2 levels contribute to mucosal damage,inflammation,and disruption of the intestinal barrier.The effects of PGE2 on macrophage activation and Maxi-Cl channel functionality,as well as its interaction with nonsteroidal anti-inflammatory drugs play crucial roles in the progression of CEAS.Understanding the balance between its protective and pro-inflammatory effects and the complex interactions within the gastrointestinal tract can shed light on potential therapeutic targets for CEAS and guide the development of novel,targeted therapies.

Transglutaminase 2 serves as a pathogenic hub gene of KRAS mutant colon cancer based on integrated analysis

BACKGROUND Colon cancer is acknowledged as one of the most common malignancies worldwide,ranking third in United States regarding incidence and mortality.Notably,approximately 40%of colon cancer cases harbor oncogenic KRAS mutations,resulting in the continuous activation of epidermal growth factor receptor signaling.AIM To investigate the key pathogenic genes in KRAS mutant colon cancer holds considerable importance.METHODS Weighted gene co-expression network analysis,in combination with additional bioinformatics analysis,were conducted to screen the key factors driving the progression of KRAS mutant colon cancer.Meanwhile,various in vitro experiments were also conducted to explore the biological function of transglutaminase 2(TGM2).RESULTS Integrated analysis demonstrated that TGM2 acted as an independent prognostic factor for progression-free survival.Immunohistochemical analysis on tissue microarrays revealed that TGM2 was associated with an elevated probability of perineural invasion in patients with KRAS mutant colon cancer.Additionally,biological roles of the key gene TGM2 was also assessed,suggesting that the downregulation of TGM2 attenuated the proliferation,invasion,and migration of the KRAS mutant colon cancer cell line.CONCLUSION This study underscores the potential significance of TGM2 in the progression of KRAS mutant colon cancer.This insight not only offers a theoretical foundation for therapeutic approaches but also highlights the need for additional clinical trials and fundamental research to support our preliminary findings.

槐杞黄颗粒辅助治疗对支气管哮喘患儿疗效及血清FGF-2、IL-33、ICAM-1水平影响

目的探究槐杞黄颗粒辅助治疗对支气管哮喘患儿疗效及血清成纤维细胞生长因子2(FGF-2)、白细胞介素-33(IL-33)、细胞间黏附分子(ICAM-1)水平的影响。方法选择2020年6月至2023年6月中江县妇幼保健院接诊的92例支气管哮喘患儿作为研究对象,根据治疗方式分为孟鲁司特钠组(n=30,孟鲁司特钠药物治疗)、槐杞黄颗粒组(n=30,槐杞黄颗粒治疗)及联合组(n=32,孟鲁司特钠+槐杞黄颗粒治疗)。比较三组疗效、肺功能及免疫血清指标IgE、IgG、IgM、外周血FGF-2、IL-33、ICAM-1表达水平、随访3个月内疾病控制情况。结果孟鲁司特钠组、槐杞黄颗粒组及联合组治疗的总有效率比较差异有统计学意义(χ^(2)=6.524,P<0.05)。三组治疗后FVC、FEV_(1)、FEV_(1)/FVC及IgG较治疗前升高,联合组均高于孟鲁司特钠组及槐杞黄颗粒组(F=8.594、11.090、6.221、8.117,P<0.05)。三组治疗后IgE、IgM、FGF-2、IL-33及ICAM-1水平较治疗前降低,联合组均低于孟鲁司特钠组及槐杞黄颗粒组(F=5.952、9.515、40.512、9.724、34.636,P<0.05)。随访3个月内,联合组喘息发作、因喘息住院、急救药物使用情况显著减少(χ^(2)=24.337、20.391、8.105,P<0.05)。结论槐杞黄颗粒与孟鲁司特钠在治疗儿童支气管哮喘方面的联合应用疗效确切,改善患儿肺功能及免疫状态,且预后较好。

Gene expression analysis of cytokines and MMPs in melatonin and rhBMP-2 enhanced bone remodeling

BACKGROUND In the medical and dental fields,there is a need for studies of new therapeutic approaches for the treatment of bone defects that cause extensive bone loss.Melatonin may be an important endogenous biological factor for bone remodeling,and growth factors may enhance the repair process.AIM To evaluate the gene expression of cytokines(IL-1β,IL-6,IL-10 and TNF-α),markers of osteoclastogenesis(RANK,RANKL and OPG)and MMPs(MMP-1,MMP-2,MMP-8 and MMP-13)from the treatment of melatonin associated with an osteogenic membrane and rhBMP-2 on the recovery of a bone injury.METHODS Sixty-four rats were used and divided into 9 experimental groups and were formed according to the treatment carried out in the region of the bone lesion,which varied between the combination of 1,10 and 100μmol/L of melatonin.Gene Expression analysis was performed using real time-PCR by reading the concentration of total RNA and reverse transcription.RESULTS There were differences between groups when compared with clot or scaffold control,and improvement with a higher concentration of melatonin or rhBMP-2.The combination melatonin(1μg)with 5μg of rhBMP-2,using the guided bone regeneration technique,demonstrated some effects,albeit mild,on bone repair of critical bone defects.CONCLUSION This indicates that the approach for administering these substances needs to be reassessed,with the goal of ensuring their direct application to the affected area.Therefore,future research must be carried out,seeking to produce materials with these ideal characteristics.

Identification of M2 macrophage-related genes for establishing a prognostic model in pancreatic cancer: FCGR3A as key gene

Background:Pancreatic ductal adenocarcinoma(PDAC)has a rich and complex tumor immune microenvironment(TIME).M2 macrophages are among the most extensively infiltrated immune cells in the TIME and are necessary for the growth and migration of cancers.However,the mechanisms and targets mediating M2 macrophage infiltration in pancreatic cancer remain elusive.Methods:The M2 macrophage infiltration score of patients was assessed using the xCell algorithm.Using weighted gene co-expression network analysis(WGCNA),module genes associated with M2 macrophages were identified,and a predictive model was designed.The variations in immunological cell patterns,cancer mutations,and enrichment pathways between the cohorts with the high-and low-risk were examined.Additionally,the expression of FCGR3A and RNASE2,as well as their association with M2 macrophages were evaluated using the HPA,TNMplot,and GEPIA2 databases and verified by tissue immunofluorescence staining.Moreover,in vitro cell experiments were conducted,where FCGR3A was knocked down in pancreatic cancer cells using siRNA to analyze its effects on M2 macrophage infiltration,tumor proliferation,and metastasis.Results:The prognosis of patients in high-risk and low-risk groups was successfully distinguished using a prognostic risk score model of M2 macrophage-related genes(p=0.024).Between the high-and low-risk cohorts,there have been notable variations in immune cell infiltration patterns,tumor mutations,and biological functions.The risk score was linked to the manifestation of prevalent immunological checkpoints,immunological scores,and stroma values(all p<0.05).In vitro experiments and tissue immunofluorescence staining revealed that FCGR3A can promote the infiltration or polarization of M2 macrophages and enhance tumor proliferation and migration.Conclusions:In this study,an M2 macrophage-related pancreatic cancer risk score model was established,and found that FCGR3A was correlated with tumor formation,metastasis,and M2 macrophage infiltration.

脂联素联合FGF-21、A-FABP在2型糖尿病的早期诊断中的应用价值分析

了解脂联素+FGF-21+A-FABP检测对于2型糖尿病早期诊断的价值。方法 选取2020.01-2023.12共计三年时间,本院住院T2DM病人及正常人群体检者300名。按照随机数表法将上述研究对象分为三组,A组:正常人群组(n=100),B组:糖尿病高危人群组(n=100),C组:糖尿病(n=100),符合糖尿病诊断标准。收集三组的基本信息,了解病情情况,并进行相关指标检测,进行对比分析。结果 糖尿病组和糖尿病高危人群组的ADPN明显低于正常人群组,而FGF-21、A-FABP明显高于正常人群组。同时,糖尿病组的ADPN明显低于糖尿病高危人群组,而FGF-21、A-FABP明显高于糖尿病高危人群组。结论 T2DM患者体内ADPN、FGF-21、A-FABP水平与病情息息相关,为患者提供涵盖这三个指标的综合检测手段,可以帮助早期诊断T2DM,以及建立起科学的治疗方案,拥有良好的临床应用价值。

BMP-2和FGF-2对小鼠骨髓间充质干细胞向成骨细胞分化的影响

目的：观察骨形成蛋白（BMP-2）和成纤维细胞生长因子2（FGF-2）对小鼠骨髓间充质干细胞（MSCs）向成骨细胞分化的影响。方法：取3-18月雄性C57BL/6J小鼠（共50只）的骨髓细胞,分离贴壁培养后,用免疫磁珠法纯化,并鉴定为MSCs后,再进行贴壁培养24h后,分别在成骨细胞诱导培养液中加入100μg/LBMP-2和0.5nmol/LFGF-2持续诱导7、14、21d后,进行碱性磷酸酶染色、碱性磷酸酶活性检测,Vonkossa染色以及茜素红染色,并用递转录荧光定量PCR法检测向成骨细胞分化的标志性基因（Runx2/cbfa1、Alp、collagen-1、osteocalcin）的表达情况。结果：BMP-2刺激组的ALP活性以及钙化结节明显高于对照组,Runx2/cbfa1,ALP,collage-1,osteocalcin的mRNA呈高表达;FGF-2刺激组的ALP活性以及钙化结节也高于对照组,Runx2/cbfa1、Alp、collagen-1、osteocalcin的mRNA表达量也高于对照组,但不如BMP-2明显。结论：BMP-2和FGF-2在不同程度上促进体外培养的小鼠MSCs向成骨细胞方向分化。

黄芪、莪术配伍对人卵巢癌HO-8910原位移植瘤组织中MMP-2、FGF-2、BCL-2表达的影响

目的：观察黄芪、莪术配伍对人卵巢癌HO-8910原位移植瘤的抑瘤效果及瘤组织中MMP-2、FGF-2、BCL-2蛋白和基因表达的影响。方法：选取已建立荧光蛋白转染的HO-8910卵巢癌原位癌动物模型,分为模型组,阳性对照组（顺铂）,黄芪组,莪术组,黄芪、莪术配伍（按照2∶1）高、中、低剂量组,监测抑瘤效果。用免疫组化和Real time-PCR检测移植瘤组织中MMP-2、FGF-2、BCL-2蛋白和基因的表达情况。结果：阳性对照组,黄芪组,黄芪、莪术配伍高剂量组,黄芪、莪术配伍低剂量组平均肿瘤质量明显小于模型组（P〈0.05）;各组MMP-2、FGF-2、BCL-2蛋白表达与模型组相比无统计学差异（P〉0.05）;黄芪组,莪术组,黄芪、莪术配伍组高、中、低剂量组肿瘤组织中的MMP-2、FGF-2、BCL-2 mRNA表达明显低于模型组（P〈0.05）。结论：黄芪、莪术配伍使用在实验周期内对人卵巢癌HO-8910具有抑制作用（P〈0.05）,其作用机制可能与下调移植瘤中MMP-2、FGF-2、BCL-2基因表达相关。

骨折三期治疗对骨生长因子FGF-2表达影响的实验研究

目的 :观察骨折三期治疗对FGF 2在骨折愈合过程中表达的影响。方法 :16 8只雄性SD大鼠随机取 6只作为正常对照组 ,其余SD大鼠双胫骨上 1/ 3段前部的中点造成 2mm× 1 5mm的缺损。术后随机平均分为模型对照组、一期治疗组和三期治疗组 ,每组于不同的时间点分别处死 6只 ,取材 ,常规组织学检查 ,免疫组化法检测标本中的FGF 2 ,应用计算机图像分析系统进行定量分析。结果 :①缺损区内血肿细胞、炎性细胞、间充质细胞、成骨细胞、新形成骨细胞FGF 2表达呈强阳性 ,成熟骨细胞阴性或微弱阳性 ;②骨折后三组FGF 2表达阳性细胞数和平均光密度值均即明显增高 ,但是 ,中药治疗组显著高于模型对照组 (P <0 0 5 ) ,三期治疗组有高于一期治疗组的趋势 ,FGF 2表达高峰出现时间中药治疗组显著早于模型组 ,三期治疗组较一期治疗组提前了 80 %。结论 :中药治疗能显著促进骨折愈合过程中FGF 2的表达 ,三期治疗明显强于一期治疗 ,因此对骨折进行分期辨证治疗是非常必要的。

初诊2型糖尿病患者短期胰岛素泵强化治疗对血浆FGF-21水平及胰岛素敏感性的影响

目的探讨短期胰岛素泵强化治疗对初诊2型糖尿病(T2DM)患者胰岛素敏感性和血浆成纤维细胞生长因子-21(FGF-21)水平的影响。方法采用自身前后对照,运用高胰岛素-正葡萄糖钳夹术评价30例新诊断T2DM患者接受2 w短期胰岛素泵(CSII)强化治疗前后胰岛素敏感性的变化,并采用酶免法测定治疗前后血浆FGF-21水平。结果 T2DM患者血浆FGF-21水平明显高于对照组〔(1.6±0.1)vs(1.1±0.4)μg/L,P<0.01〕。T2DM患者经胰岛素泵强化治疗2 w后,血浆FGF-21水平较治疗前明显下降〔(1.6±0.1)vs(1.3±0.1)μg/L,P<0.05〕。多元逐步回归分析表明,WHR、2 h PBG、SBP及FFA是影响血浆FGF-21水平的独立相关因素(YFGF-21=0.625XWHR+0.034XPBG+0.003XSBP-0.419XFFA+0.626)。结论短期胰岛素泵强化治疗可明显降低T2DM患者血浆FGF-21水平,血浆FGF-21水平与代谢紊乱有关,并可能参与了T2DM的发生和发展。

糖尿病肾病患者血清FGF-2、TGF-β及CD4^+、CD8^+T细胞的数量变化

糖尿病肾病（diabetic nephropathy，DN）是糖尿疖全身微血管合并症之一，最终可发展成终末期肾病也是糖尿病最主要的死亡原因之一[1-2]。糖尿病肾病在临床上分为5期，一旦发病，缺乏有效的方法空制其发展，大多数患者转为慢性肾功能衰竭，威胁毒患者的生命。

成纤维细胞生长因子(FGF)受体-2参与FGF-21介导的糖代谢活性

最近发现,FGF-21具有很强的调节血糖和血脂的作用,已经成为糖尿病研究领域的新热点,但是其功能受体和作用机制还不清楚.前期结果表明,FGF-21促进3T3L1脂肪细胞代谢葡萄糖,对前脂肪细胞无作用,说明脂肪细胞表达FGF-21功能受体.以3T3L1脂肪细胞为靶标,旨在寻找FGF-21的功能受体.结果表明,FGF-21可与3T3L1脂肪细胞膜蛋白形成FGF-21/受体复合物,免疫检测结果发现,FGF-21/受体复合物中含有FGF受体-2(FGFR-2).为明确FGF-21/FGFR-2的特异性关系,系统研究了FGFR-2对FGF-21刺激后的酪氨酸磷酸化反应.结果表明,虽然前脂肪细胞和脂肪细胞均表达FGFR-2,但是FGF-21只诱导脂肪细胞中表达的FGFR-2磷酸化,对前脂肪细胞表达的FGFR-2无作用,与葡萄糖吸收试验相符.FGF-21不仅可使原位表达的FGFR-2磷酸化,还可使异位表达的FGFR-2磷酸化.克隆后测序分析结果表明,FGFR-2Ⅲc是3T3L1脂肪细胞表达的主要FGFR-2类型.这些结果提示,FGFR-2Ⅲc是FGF-21的功能受体,参与FGF-21在脂肪细胞介导的糖代谢活性.此外,系统分析了FGFR-2在3T3L1分化过程中的差异表达,为FGF-21在前脂肪细胞和脂肪细胞中的功能差异提供了依据.

FGF-2和VEGF在慢性应激抑郁大鼠前额叶皮层表达

目的探讨慢性应激抑郁大鼠前额叶皮层成纤维细胞生长因子-2(fibroblast growth factors-2,FGF-2)和血管内皮生长因子(vascular endothelial growth factor,VEGF)表达变化,分析其在抑郁症病理生理学中的意义。方法 20只体质量180～220 g雄性SD大鼠分为正常对照组(对照组)、慢性应激抑郁模型组(抑郁组),每组10只。利用21 d慢性不可预见性应激刺激(chronic unprediectable mild stress,CUMS)制造抑郁症模型。21 d后,采用敞箱实验(open-field behavior)、糖水消耗实验、摄食量和体质量增量等检测指标评定大鼠行为学改变并检测模型是否成功建立。应用Western blot法检测FGF-2和VEGF蛋白表达情况;荧光实时定量PCR技术检测目的基因mRNA表达情况。结果 Western blot检测结果显示:抑郁组SD大鼠前额叶皮质层FGF-2(1.23±0.06)、VEGF蛋白表达(1.15±0.27)显著低于对照组(P<0.05);rt-q PCR结果显示:与对照组相比,抑郁组SD大鼠前额叶皮质层FGF-2的2-△△Ct值为0.47、VEGF的2-△△Ct值为0.51,提示二者mRNA表达与对照组相比显著下调(P<0.05)。结论 FGF-2与VEGF基因在前额叶中的表达下调可能在抑郁症病理机制中发挥一定作用。