Interferon beta(IFN-β) treatment exerts potential neuroprotective effects through neurotrophic factors and novel neurotensin/neurotensin high affinity receptor 1 pathway

Multiple sclerosis（MS）is a chronic autoimmune disease of the central nervous system（CNS）characterized by coexisting processes of inflammation,demyelination,axonal neurodegeneration,and gliosis.It is the most common disabling neurological disease in young adulthood.

Myeloid neoplasm with eosinophilia and rearrangement of plateletderived growth factor receptor beta gene in children: Two case reports

BACKGROUND Myeloid neoplasm(MN)with eosinophilia and rearrangement of platelet-derived growth factor receptor beta(PDGFRB)shows a good therapeutic response to imatinib in adults.MN is rarely found in children,and the efficacy of imatinib on pediatric patients remain unclear.CASE SUMMARY We report 2 pediatric cases diagnosed with MN with eosinophilia and PDGFRB rearrangement who were treated with imatinib.Case 1 was a 1-year-old girl admitted to the hospital because of“abdominal distension with hyperleukocytosis for 3 mo”.She had leukocytosis,anemia,and eosinophilia(the absolute eosinophil count(AEC)was 8960/μL),and her fluorescence in situ hybridization(FISH)test revealed that PDGFRB rearrangement was detected in 70%of 500 interphase cells.Case 2 was a 2-year-old girl admitted to the hospital because of“recurrent fever and rashes for 1 mo”.Her blood cell count showed an AEC of 3540/μL.The FISH test revealed that PDGFRB rearrangement was detected in 71%of 500 interphase cells.Both patients were diagnosed as MN with eosinophilia and PDGFRB rearrangement.Imatinib was added into their treatment regimen.As expected,complete hematologic remission was achieved after 1 mo of treatment,and symptoms disappeared.CONCLUSION Although MN with eosinophilia and PDGFRB rearrangement usually occurs in adults,it can be found in children.The therapeutic benefits of imatinib in these 2 pediatric patients were consistent with its reported effects in adult patients.

Expression patterns of transforming growth factor-beta and its receptors in gastric mucosa of patients with refractory gastric ulcer

AIM: Transforming growth factor-β (TGF-β) plays a regulatory role in tissue repair. In a previous study, we found that TGF-β and its receptors were expressed in gastric mucosa of patients with well-healed gastric ulcers, as demonstrated by immunohistochemistry. To further characterize the role of TGF-β and its receptors in repairing gastric ulcers, we investigated the expression patterns of TGF-β and its receptors in gastric mucosa by in situ hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR).METHODS: Seventy-four patients with endoscopically proven gastric ulcers were eligible for participation in this study. All patients had routine biopsies on initial endoscopy and were then treated for 12 wk with an H2 blocker. Repeat endoscopy was then performed. There were 8 patients with poorly healed ulcers, and biopsies were taken from the margin of the residual ulcers. These tissue samples, along with biopsy of gastric mucosa near the original ulcers from 8 randomly selected patients with well-healed ulcers were examined for TGF-β and TGF-β receptor Ⅱ mRNA by RT-PCR and in situ hybridization, as well as immunohistochemistry.RESULTS: TGF-β and TGF-β receptor Ⅱ were strongly expressed in tissues from patients with well-healed ulcers.Four of the 8 patients with poor healing had low or absent expression of TGF-β or TGF-β receptor Ⅱ mRNA. All cases positive by RT-PCR assay were confirmed by in situ hybridization as well as immunohistochemistry.CONCLUSION: It is suggested that TGF-β and its receptors are important for gastric ulcer healing. These results may have implications for further investigation of the healing process and in predicting response to therapy.

Transforming growth factor-β and toll-like receptor-4 polymorphisms are not associated with fibrosis in haemochromatosis

AIM:To investigate the role of genetic polymorphisms in the progression of hepatic fibrosis in hereditary haemochromatosis.METHODS:A cohort of 245 well-characterised C282Y homozygous patients with haemochromatosis was studied,with all subjects having liver biopsy data and DNA available for testing.This study assessed the association of eight single nucleotide polymorphisms(SNPs)in a total of six genes including toll-like receptor 4(TLR4),transforming growth factor-beta(TGF-β),oxoguanine DNA glycosylase,monocyte chemoattractant protein 1,chemokine C-C motif receptor 2 and interleukin-10 with liver disease severity.Genotyping was performed using high resolution melt analysis and sequencing.The results were analysed in relation to the stage of hepatic fibrosis in multivariate analysis incorporating other cofactors including alcohol consumption and hepatic iron concentration.RESULTS:There were significant associations between the cofactors of male gender(P=0.0001),increasing age(P=0.006),alcohol consumption(P=0.0001),steatosis(P=0.03),hepatic iron concentration(P<0.0001)and the presence of hepatic fibrosis.Of the candidate gene polymorphisms studied,none showed a significant association with hepatic fibrosis in univariate or multivariate analysis incorporating cofactors.We also specifically studied patients with hepatic iron loading above threshold levels for cirrhosis and compared the genetic polymorphisms between those with no fibrosis vs cirrhosis however there was no significant effect from any of the candidate genes studied.Importantly,in this large,well characterised cohort of patients there was no association between SNPs for TGF-βor TLR4and the presence of fibrosis,cirrhosis or increasing fibrosis stage in multivariate analysis.CONCLUSION:In our large,well characterised group of haemochromatosis subjects we did not demonstrate any relationship between candidate gene polymorphisms and hepatic fibrosis or cirrhosis.

Inhibiting 5-hydroxytryptamine receptor 3 alleviates pathological changes of a mouse model of Alzheimer's disease

Extracellular amyloid beta(Aβ) plaques are main pathological feature of Alzheimer’s disease.However,the specific type of neuro ns that produce Aβ peptides in the initial stage of Alzheimer’s disease are unknown.In this study,we found that 5-hydroxytryptamin receptor 3A subunit(HTR3A) was highly expressed in the brain tissue of transgenic amyloid precursor protein and presenilin-1 mice(an Alzheimer’s disease model) and patients with Alzheimer’s disease.To investigate whether HTR3A-positive interneurons are associated with the production of Aβ plaques,we performed double immunostaining and found that HTR3A-positive interneurons were clustered around Aβ plaques in the mouse model.Some amyloid precursor protein-positive or β-site amyloid precursor protein cleaving enzyme-1-positive neurites near Aβ plaques were co-localized with HTR3A interneurons.These results suggest that HTR3A-positive interneurons may partially contribute to the generation of Aβ peptides.We treated 5.0-5.5-month-old model mice with tro pisetron,a HTR3 antagonist,for 8 consecutive weeks.We found that the cognitive deficit of mice was partially reversed,Aβ plaques and neuroinflammation we re remarkably reduced,the expression of HTR3 was remarkably decreased and the calcineurin/nuclear factor of activated T-cell 4 signaling pathway was inhibited in treated model mice.These findings suggest that HTR3A interneurons partly contribute to generation of Aβ peptide at the initial stage of Alzheimer’s disease and inhibiting HTR3 partly reve rses the pathological changes of Alzheimer’s disease.

Synergistic effect of interleukin-10-receptor variants in a case of early-onset ulcerative colitis

AIM:To investigated the molecular cause of very early-onset ulcerative colitis(UC)in an 18-mo-old affected child.METHODS:We analysed the interleukin-10(IL10)receptor genes at the DNA and RNA level in the proband and his relatives.Beta catenin and tumor necrosis factor-α(TNFα)receptors were analysed in the proteins extracted from peripheral blood cells of the proband,his relatives and familial adenomatous polyposis(FAP)and PTEN hamartoma tumor syndrome(PHTS)patients.Samples were also collected from the proband’s inflamed colorectal mucosa and compared to healthy and tumour mucosa collected from a FAP patient and patients affected by sporadic colorectal cancer(CRC).Finally,we examined mesalazine and azathioprine effects on primary fibroblasts stabilised from UC and FAP patients.RESULTS:Our patient was a compound heterozygote for the IL10RB E47K polymorphism,inherited from his father,and for a novel point mutation within the IL10RA promoter(the-413G->T),inherited from his mother.Beta catenin and tumour necrosis factorαreceptors-Ⅰ(TNFRI)protein were both over-expressed in peripheral blood cells of the proband’s relatives more than the proband.However,TNFRII was over-expressed only in the proband.Finally,both TNFα-receptors were shown to be under-expressed in the inflamed colon mucosa and colorectal cancer tissue compared to healthy colon mucosa.Consistent with this observation,mesalazine and azathioprine induced,in primary fibroblasts,IL10RB and TNFRII over-expression and TNFRI and TNFαunder-expression.We suggest thatβ-catenin and TNFRI protein expression in peripheral blood cells could represent molecular markers of sub-clinical disease in apparently healthy relatives of patients with early-onset UC.CONCLUSION:A synergistic effect of several variant alleles of the IL10 receptor genes,inherited in a Mende-lian manner,is involved in UC onset in this young child.

Role of Toll-like receptor 4 in inflammatory reactions of hippocampal neurons

Lipopolysaccharide stimulates Toll-like receptor 4 on immune cells to produce immune mediators. Toll-like receptor 4 is also expressed by non-immune cells, which can be stimulated by lipopolysaccharide. However, whether Toll-like receptor 4 is expressed by primary cultured hippocampal neurons and its specific role in lipopolysaccharide-induced neuroinflammation is currently undefined, in this study, Toll-like receptor 4 antibody blocking was used to analyze the Toll-like receptor 4 signaling pathway and changes in inflammation of lipopolysaccharide stimulated hippocampal neurons. Immunofluorescence showed that Toll-like receptor 4 protein was mainly located in the membrane of hippocampal neurons. Quantitative reverse transcription-PCR and western blot assay showed that after stimulation of lipopolysaccharide, the mRNA and protein levels of Toll-like receptor 4 and the mRNA levels of interleukin-ll3 and tumor necrosis factor-（] were significantly increased. In addition, there was increased phosphorylation and degradation of kappa B a inhibitor in the cytosol and increased nuclear factor-KB p65 expression in the nuclei. Pretreatment with Toll-like receptor 4 antibody could almost completely block this increase. These experimental findings indicate that lipopolysaccharide participates in neuroinflammation by stimulating Toll-like receptor 4/nuclear factor-KB pathway in hippocampal neurons, which may be both ＂passive victims＂ and ＂activators＂ of neuroinflammation.

Role of activin receptor-like kinase 1 in vascular development and cerebrovascular diseases

Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,and its dynamic changes are closely related to the proliferation of endothelial cells,the recruitment of pericytes to blood vessels,and functional differentiation during embryonic vascular development.The pathophysiology of many cerebrovascular diseases is today understood as a disorder of endothelial cell function and an imbalance in the proportion of vascular cells.Indeed,mutations in ALK1 and its co-receptor endoglin are major genetic risk factors for vascular arteriovenous malformation.Many studies have shown that ALK1 is closely related to the development of cerebral aneurysms,arteriovenous malformations,and cerebral atherosclerosis.In this review,we describe the various roles of ALK1 in the regulation of angiogenesis and in the maintenance of cerebral vascular homeostasis,and we discuss its relationship to functional dysregulation in cerebrovascular diseases.This review should provide new perspectives for basic research on cerebrovascular diseases and offer more effective targets and strategies for clinical diagnosis,treatment,and prevention.

Down-regulation of peroxisome proliferator-activated receptor γ coactivator-1α expression in fatty acid-induced pancreatic betacell apoptosis involves nuclear factor-κB pathway

Background Pancreatic beta-cell apoptosis induced by lipotoxicity, to a large extent, contributes to the progression of type 2 diabetes. To investigate the mechanism of free fatty acid induced apoptosis, we aimed to study the effects of palmitic acid （PA） on the apoptosis and peroxisome proliferator-activated receptor y coactivator-1α （PGC-1α） expression in βTC3 cells as well as the possible role of nuclear factor-KB （NF-KB） in this process. Methods Hoechst 33258 was used to detect βTC3 cell apoptosis, which was induced by PA stimulation for 12 hours. PGC-1α expression was analyzed by reverse transcription polymerase chain reaction, IκB kinase β （IKKβ）, IκBα NF-KB-inducing kinase （NIK） and ReI-B expressions were analyzed by Western blotting. MGβ2 was employed to block the endogenous IκBαdegradation before PA administration, and then its effect on PA-inducing cell apoptosis and PGC-1α mRNA expression was analyzed. Results Significant increased cell apoptosis was found at the concentration of 0.5 mmol/L and 1.0 mmol/L PA administration. PA （0.5 mmol/L） could extensively reduced the expression of PGC-1α mRNA. After exposing βTC3 cells to 0.5 mmol/L PA for different time periods （0, 4, 6, 8, 10 and 12 hours）, IKKβ protein expression increased while IκBα NIK and ReI-B protein expression declined in a time-dependent manner. Pretreatment with MGβ2 to inhibit the degradation of IκBα partially prevented the down-regulation of PGC-1α mRNA expression after 12-hour PA treatment in accordance with the decrease of PA induced apoptosis. Conclusions NF-KB canonical pathway was activated in PA-mediated βTC3 cell apoptosis, whereas non-canonical pathway was inhibited. Reduced PGC-1α expression by PA in βTC3 cells could involve the activation of canonical NF-KB pathway, so as to deteriorate the PA induced apoptosis.

Effect of substance P on gene expression of transforming growth factor β-1 and its receptors in rat's fibroblasts

Objective: To investigate the effect of substance P (SP) on gene expression of transforming growth factor β-1 (TGFβ-1), transforming growth factor receptor-1 (TGFR-1) and transforming growth factor receptor-2 (TGFR-2) in fibroblasts cultured in vitro from rat’s granulation tissues. Methods: The fibroblasts from the granulation tissues in the skeletal muscle of rat’s hind limbs injured by formaldehyde were cultured in vitro. When different concentrations (10 -9-10 -5 mol/L) of SP were added into the culture medium, the changes of gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the cultured fibroblasts were observed with reverse transcription polymerase chain reaction at different intervals (0, 3, 6, 12 and 24 hours after incubation). Results: The gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the fibroblasts cultured from rat’s granulation tissues was up-regulated by SP. The peak level of the mRNA expression was found at 10 -8 mol/L SP and the up-regulation effect was not found at 10 -5 mol/L and 10 -6 mol/L. The peak levels of gene expression of TGFβ-1, TGFR-1 and TGFR-2 in the fibroblasts treated with SP were achieved at 6 and 12 hours, respectively. Conclusions: SP has up-regulation effect on the gene expression of TGFβ-1, TGFR-1 and TGFR-2 in fibroblasts from rat’s granulation tissues in vitro, and the effect is related to different stimulating concentrations of SP. It may be concerned with proliferation and differentiation of fibroblasts and formation of scar tissues during wound healing.

伴有微卫星不稳定性的散发性结直肠癌转化生长因子 -betaⅡ型受体基因突变的研究(英文)

目的 :为了探讨在散发性结直肠癌的发生过程中转化生长因子βⅡ型受体基因 (RII)的突变与微卫星不稳定性 (RER)间的关系。方法 :我们应用PCR -SSCP -银染方法检测了 5 0例散发性结直肠癌中的RER状态及RII基因突变情况 (近端结肠 19例 ,远端 31例 )。结果 :RER +的共有 13例 (8例在近端 ,5例在远端 ) ,RII基因突变的共有 5例。所有 5例RII基因突变者均同时伴有RER +,而所有RER -病例均无RII基因突变。其中 4例RII基因突变者位于回盲部肿瘤中。结论 :这些数据提示在散发性结直肠癌中 ,尤其是在回盲部肿瘤中 ,TGF - βRII基因的A10 重复序列是微卫星不稳定性的靶点 。

miR-10b靶向TGFBR1/SMAD3通路对特发性矮小症的软骨细胞增殖、肥大的影响机制

目的研究miR-10b对特发性矮小症(ISS)的影响及作用机制。方法收集ISS患儿(ISS组)和体检健康儿童(健康对照组)各54例,qPCR检验血清miR-10b表达量,分析ISS组患儿血清miR-10b表达与患儿临床资料的关系。采用miR-10b inhibitor、si-TGFBR1及各自阴性对照转染C28/I2细胞,利用CCK-8实验检测C28/I2细胞增殖能力,Western blot检测侏儒相关转录因子2(RUNX2)、X型胶原α1链(COL10A1)、转化生长因子β受体1(TGFBR1)、SMAD3、pSMAD3蛋白表达量。在StarBase数据库筛选miR-10b靶点,利用双萤光素酶报告基因实验验证miR-10b与TGFBR1的靶向关系。结果ISS组血清miR-10b表达量高于健康对照组,且miR-10b表达越高,患儿的身高、IGF-1、骨特异性碱性磷酸酶的下降越明显(P<0.05)。与NC组相比,miR-10b inhibitor组细胞增殖能力升高,RUNX2、COL10A1、TGFBR1、pSMAD3蛋白表达上调(P<0.05);StarBase数据库提示miR-10b存在TGFBR1的结合位点,双萤光素酶报告基因实验证实两者结合。与si-NC相比,si-TGFBR1组TGFBR1表达量下调,细胞增殖能力下降(P<0.05)。结论miR-10b通过靶向TGFBR1/SMAD3通路在特发性矮小症中抑制软骨细胞的增殖、肥大。

Activation of PPAR-γ Inhibits Differentiation of Rat Osteoblasts by Reducing Expression of Connective Tissue Growth Factor

Long-term treatment with an agonist of peroxisome proliferator-activated receptor （PPAR）-γ is associated with bone fractures in the clinical practice. However, the mechanisms underlying the frac- tures are not fully understood. This study was aimed to examine the effect of rosiglitazone （an agonist of PPAR-T） of different doses on the proliferation, differentiation, and transforming growth factor beta 1 （TGF-131）-induced expression of connective tissue growth factor （CTGF） in primary rat osteoblasts in vitro. Osteoblasts were isolated from newly born SD rats and treated with different doses of rosiglita- zone （0-20 gmol/L）. The proliferation and differentiation of osteoblasts were measured by MTT assay and NPP assay, respectively. The expression of CTGF was determined by RT-PCR and Western blotting. The results showed that most isolated osteoblasts displayed strong alkaline phosphatase （ALP） activity and treatment with different doses of rosiglitazone did not affect their proliferation, but significantly in- hibited the differentiation of osteoblasts in a dose-dependent manner. Moreover, treatment with different doses of rosiglitazone significantly reduced the TGF-131-induced CTGF mRNA transcription and protein expression in a dose-dependent manner in rat osteoblasts. It was concluded that the activation of PPAR-y may inhibit the differentiation of osteoblasts by reducing the TGF-131-induced CTGF expres- sion in vitro.

PDGFR-β/TGF-β/Smad2/3信号通路调控阿尔茨海默病血脑屏障完整性和学习记忆能力的分子机制研究

目的:分析PDGFR-β/TGF-β/Smad2/3信号通路调控阿尔茨海默病(AD)血脑屏障(BBB)完整性和学习记忆能力的分子机制。方法:利用APP/PS1转基因AD小鼠模型,通过水迷路及觅食试验分析学习记忆能力;荧光免疫组织化学法检测海马区血管周细胞增殖率(ki67/desmin)、周细胞覆盖率(desmin/lectin);Western blot检测海马区血管周细胞TGF-βR1及其下游信号通路分子、紧密连接(TJs)蛋白的表达水平。经过外源性PDGF-BB脑室内注射和/或TGF-βR1激酶抑制剂SB431542腹腔内注射预处理后,分别进行上述分析。构建AD体外BBB模型,经过PDGF-BB和/或SB431542作用后,进行异硫氰酸荧光素-牛血清白蛋白(FITC-BSA)渗透性和跨细胞电阻检测。结果:与对照组相比,APP/PS1小鼠经过虚拟平台次数明显减少,达到终点所需时间明显延长(水迷路训练试验),投食区正确选择率明显下降(觅食训练试验);海马区desmin/lectin阳性细胞比例明显下降;TGF-βR1、p-Smad2、p-Smad3蛋白表达水平明显升高;TJs蛋白表达水平明显下降。外源性PDGF-BB可使APP/PS1小鼠经过虚拟平台次数明显增加、达到终点所需时间明显缩短(水迷路正式试验第28天)、投食区正确选择率明显提高(觅食正式试验第28天);海马区desmin/lectin阳性细胞比例明显增加;使TGF-βR1、p-Smad2、p-Smad3蛋白表达水平明显升高;使TJs蛋白表达水平明显升高。SB431542则可部分抑制上述作用。体外试验证明:外源性PDGF-BB可明显降低AD模型组的最终渗透系数,提高24 h时相对TEER值;SB431542则可部分抑制上述作用。结论:PDGFR-β/TGF-β/Smad2/3信号通路可通过诱导周细胞分化、覆盖,提高内皮细胞TJs的表达,调控AD血脑屏障完整性,以促进学习记忆能力恢复。

遗传调控下免疫相关血浆蛋白对帕金森病的效应

目的 探讨免疫相关血浆蛋白与帕金森病(Parkinson's disease, PD)的联系。方法 通过对4907种免疫相关血浆蛋白的全基因组关联研究数据进行分析,评估血浆蛋白对PD风险的直接影响。研究还利用单细胞核RNA测序数据进行蛋白表达分析。结果 4种免疫相关蛋白质脑源性多巴胺营养因子(cerebral dopamine neurotrophic factor, CDNF)、组织蛋白酶B(cathepsin B,CTSB)、免疫球蛋白G Fc受体2a(FCGR2A)、血红蛋白β亚基(HBB)与PD风险存在潜在联系;其中,CDNF、CTSB、HBB表达增高有助于降低PD风险(OR=0.871,95%CI:0.779~0.973,P=0.015;OR=0.835,95%CI:0.758~0.920,P=0.001;OR=0.735,95%CI:0.631~0.857,P=0.001),而FCGR2A表达增高与PD风险增高相关(OR=1.137,95%CI:1.058~1.223,P=0.001)。单细胞测序分析蛋白表达及其在脑中不同细胞类型中分布,CDNF、CTSB在大脑的多种细胞中大量表达;FCGR2A主要在大脑小胶质细胞中表达;HBB在大脑中几乎不表达。结论 研究揭示了CDNF、CTSB、FCGR2A及HBB 4种蛋白质与PD风险的潜在关联,强调了PD遗传风险变异通过调节这些免疫相关蛋白表达来影响PD发生。此外,单细胞表达数据揭示相关免疫蛋白在大脑的表达模式。

大白鼠肝脏Beta肾上腺能受体随增龄的变化

本文测定了3、12和18月龄大白鼠在低温(4—6℃)和室温(20—25℃)条件下肝脏β受体数目,亲和力及cAMP(环一磷酸腺苷)含量。结果显示随增龄β受体数目增加,亲和力下降,cAMP含量减少;β受体数目在低温组分别比相应的室温组为高。表明随增龄大白鼠肝脏β受体质量下降,非寒战产热减少。

激活蛋白-1在骨代谢相关信号通路中的作用研究进展

激活蛋白-1(AP-1)是调节细胞增殖、分化和凋亡的关键转录因子。AP-1可通过调控成骨细胞和破骨细胞的增殖、分化和凋亡,参与骨骼生长的生理及病理过程。近年来研究发现,AP-1作为核因子-κB受体活化因子配体/核因子-κB受体活化因子信号通路的重要转录因子,主要通过核因子-κB和Jun氨基末端激酶、细胞外信号调节激酶和p38信号通路调节破骨细胞形成,参与骨代谢过程,其与骨质疏松、骨肿瘤等代谢性骨疾病的发生发展相关,这为代谢性骨疾病提供了新的治疗靶点。本文就AP-1在骨代谢相关信号通路的作用研究进展作一综述。

高亲和PDGFβR多肽修饰的截短型TβRⅡ的可溶性表达条件优化

目的 探讨高亲和血小板衍生生长因子受体(Platelet derived growth factor-β receptor, PDGFβR)多肽ZPDGFβR修饰的截短型转化生长因子β Ⅱ型受体(Truncated transforming growth factor-β receptor type Ⅱ,tTβRⅡ)的重组蛋白(Z-tTβRⅡ)最佳可溶性表达的条件。方法 将含有原核表达质粒pET28/Z-tTβRⅡ的阳性大肠杆菌(Rossta/pET28-Z-tTβRⅡ)置于LB培养基中培养,接着加入不同浓度的IPTG,使其终浓度为0.2、0.4、0.6、0.8 mmo/L,应用SDS-PAGE技术分析在不同温度时间下(16℃、18 h, 20℃、20 h和37℃、6 h)重组蛋白Z-tTβRⅡ的可溶性表达。结果 Z-tTβRⅡ在16℃、18 h下,0.8 mmol/L IPTG时,诱导后全菌表达约22%和诱导后上清表达约13%;Z-tTβRⅡ在20℃、20 h下,诱导后全菌0.8 mmol/L IPTG时表达约23%,诱导后上清0.2 mmol/L IPTG时表达约14%;Z-tTβRⅡ在37℃、6 h下,0.2 mmol/L IPTG时,诱导后全菌约30%和诱导后上清约20%。结论 Z-tTβRⅡ蛋白最佳表达条件为37℃、6 h、IPTG浓度为0.2 mmo/L。

糖耐康改善KKAy小鼠Toll样受体4通路及葡萄糖转运子4的作用机制研究

目的:观察中药糖耐康对KKAy小鼠血糖、体质量及Toll样受体4(TLR4)通路中白细胞介素6(IL-6)、核因子κB激酶β抑制物(IKKβ)及葡萄糖转运蛋白4(GluT4)影响,分析其改善2型糖尿病的可能作用机制。方法:选取无特定病原体(SPF)级自发性2型糖尿病动物KKAy小鼠及C57BL/6J小鼠,给予高脂饲料喂养,连续8周给药,评价血糖及体质量。并采用半定量酶联免疫吸附测定(ELISA)检测血浆中IL-6及GluT4的水平。蛋白质印迹法检测肝组织TLR4蛋白表达、肌肉组织IKKβ蛋白表达水平。结果:中药糖耐康高剂量组,能够显著降低血糖水平(P<0.05),具有一定抑制体质量增长的趋势。同时能够下调IL-6水平,上调GluT4的水平(P<0.01,P<0.05),抑制TLR4及IKKβ蛋白表达(P<0.01,P<0.05)。结论:糖耐康可能通过下调脂多糖(LPS)介导的TLR4通路中关键炎症介质的水平,减轻低度炎症反应,在一定程度上抑制炎症级联效应,同时提高胰岛素信号转导受体后途径中GluT4的转位及活性水平,共同达到改善胰岛素抵抗状态的作用。

TGFBR基因多态性与主动脉夹层的相关性研究

目的评价转化生长因子-β受体(TGFBR)基因多态性与主动脉夹层发病风险的关联。方法检索中国知网中国期刊全文数据库、万方数据库、维普数据库、美国国立医学图书馆(PubMed)、荷兰医学文摘(EMBASE)、Cochrane Library数据库,获取与TGFBR与主动脉夹层易感性的文献,检索时间截止2022年3月1日。采用RavMan5.3软件进行meta分析。结果纳入5篇文献,包括1277例病例和2019例对照。TGFBR1(rs1626340)位点等位基因模型(A/G)meta分析结果显示A等位基因更容易导致主动脉夹层疾病的发生(OR=1.25,95%CI:1.11~1.42,P=0.0004);显性基因模型(AG+AA/GG)meta分析结果显示,具有AG+AA基因的人群可能更易发生主动脉夹层(OR=1.45,95%CI:1.09~1.93,P=0.01);隐性基因模型(AA/AG+GG)的meta分析结果显示,AG+GG基因与主动脉夹层患病风险无相关性(OR=0.93,95%CI:0.66~1.32,P=0.70)。TGFBR2(rs4522809)位点等位基因模型(OR=1.01,95%CI:0.75~1.36,P=0.95)、显性基因模型(OR=1.11,95%CI:0.84~1.46,P=0.46)、隐性基因模型(OR=1.15,95%CI:0.81~1.64,P=0.44)的meta分析结果显示,TGFBR2(rs4522809)位点多态性与主动脉夹层发病风险无显著相关性(P>0.05)。结论TGFBR1基因rs1626340位点多态性与主动脉夹层疾病显著相关。