Silencing of Jumonji domain-containing 1C inhibits the osteogenic differentiation of bone marrow mesenchymal stem cells via nuclear factor-κB signaling

BACKGROUND Osteoporosis is a common metabolic bone disorder induced by an imbalance between osteoclastic activity and osteogenic activity.During osteoporosis,bone mesenchymal stem cells(BMSCs)exhibit an increased ability to differentiate into adipocytes and a decreased ability to differentiate into osteoblasts,resulting in bone loss.Jumonji domain-containing 1C(JMJD1C)has been demonstrated to suppress osteoclastogenesis.AIM To examine the effect of JMJD1C on the osteogenesis of BMSCs and the potential underlying mechanism.METHODS BMSCs were isolated from mouse bone marrow tissues.Oil Red O staining,Alizarin red staining,alkaline phosphatase staining and the expression of adipo-genic and osteogenic-associated genes were assessed to determine the differen-tiation of BMSCs.Bone marrow-derived macrophages(BMMs)were incubated with receptor activator of nuclear factor-kappaΒligand to induce osteoclast differentiation,and osteoclast differen-tiation was confirmed by tartrate-resistant acid phosphatase staining.Other related genes were measured via reverse transcription coupled to the quantitative polymerase chain reaction and western blotting.Enzyme-linked immunosorbent assays were used to measure the levels of inflammatory cytokines,including tumor necrosis factor alpha,interleukin-6 and interleukin-1 beta.RESULTS The osteogenic and adipogenic differentiation potential of BMSCs isolated from mouse bone marrow samples was evaluated.JMJD1C mRNA and protein expression was upregulated in BMSCs after osteoblast induction,while p-nuclear factor-κB(NF-κB)and inflammatory cytokines were not significantly altered.Knockdown of JMJD1C repressed osteogenic differentiation and enhanced NF-κB activation and inflammatory cytokine release in BMSCs.Moreover,JMJD1C expression decreased during BMM osteoclast differentiation.CONCLUSION The JMJD1C/NF-κB signaling pathway is potentially involved in BMSC osteogenic differentiation and may play vital roles in the pathogenesis of osteoporosis.

Growth differentiation factor-15 serum concentrations reflect disease severity and anemia in patients with inflammatory bowel disease

BACKGROUND Population of patients with inflammatory bowel disease(IBD)is burdened by various extraintestinal manifestations which substantially contribute to greater morbidity and mortality.Growth-differentiation factor-15(GDF-15)is often overexpressed under stress conditions,such as inflammation,malignancies,heart failure,myocardial ischemia,and many others.AIM To explore the association between GDF-15 and IBD as serum concentrations of GDF-15 were shown to be an independent predictor of poor outcomes in multiple diseases.An additional aim was to determine possible associations between GDF-15 and multiple clinical,anthropometric and laboratory parameters in patients with IBD.METHODS This cross-sectional study included 90 adult patients diagnosed with IBD,encompassing both Crohn’s disease(CD)and ulcerative colitis(UC),and 67 healthy age-and sex-matched controls.All patients underwent an extensive workup,including colonoscopy with subsequent histopathological analysis.Disease activity was assessed by two independent gastroenterology consultants specialized in IBD,employing well-established clinical and endoscopic scoring systems.GDF-15 serum concentrations were determined following an overnight fasting,using electrochemiluminescence immunoassay.RESULTS In patients with IBD,serum GDF-15 concentrations were significantly higher in comparison to the healthy controls[800(512-1154)pg/mL vs 412(407-424)pg/mL,P<0.001],whereas no difference in GDF-15 was found between patients with CD and UC[807(554-1451)pg/mL vs 790(509-956)pg/mL,P=0.324].Moreover,multiple linear regression analysis showed that GDF-15 levels predict CD and UC severity independent of age,sex,and C-reactive protein levels(P=0.016 and P=0.049,respectively).Finally,an association between GDF-15 and indices of anemia was established.Specifically,negative correlations were found between GDF-15 and serum iron levels(r=-0.248,P=0.021),as well as GDF-15 and hemoglobin(r=-0.351,P=0.021).Accordingly,in comparison to IBD patients with normal hemoglobin levels,GDF-15 serum levels were higher in patients with anemia(1256(502-2100)pg/mL vs 444(412-795)pg/mL,P<0.001).CONCLUSION For the first time,we demonstrated that serum concentrations of GDF-15 are elevated in patients with IBD in comparison to healthy controls,and the results imply that GDF-15 might be involved in IBD pathophysiology.Yet,it remains elusive whether GDF-15 could serve as a prognostic indicator in these patients.

Thymoquinone affects hypoxia-inducible factor-1αexpression in pancreatic cancer cells via HSP90 and PI3K/AKT/mTOR pathways

BACKGROUND Pancreatic cancer(PC)is associated with some of the worst prognoses of all major cancers.Thymoquinone(TQ)has a long history in traditional medical practice and is known for its anti-cancer,anti-inflammatory,anti-fibrosis and antioxidant pharmacological activities.Recent studies on hypoxia-inducible factor-1α(HIF-1α)and PC have shown that HIF-1αaffects the occurrence and development of PC in many aspects.In addition,TQ could inhibit the development of renal cancer by decreasing the expression of HIF-1α.Therefore,we speculate whether TQ affects HIF-1αexpression in PC cells and explore the mechanism.AIM To elucidate the effect of TQ in PC cells and the regulatory mechanism of HIF-1αexpression.METHODS Cell counting kit-8 assay,Transwell assay and flow cytometry were performed to detect the effects of TQ on the proliferative activity,migration and invasion ability and apoptosis of PANC-1 cells and normal pancreatic duct epithelial(hTERTHPNE)cells.Quantitative real-time polymerase chain reaction and western blot assay were performed to detect the expression of HIF-1αmRNA and protein in PC cells.The effects of TQ on the HIF-1αprotein initial expression pathway and ubiquitination degradation in PANC-1 cells were examined by western blot assay and co-immunoprecipitation.RESULTS TQ significantly inhibited proliferative activity,migration,and invasion ability and promoted apoptosis of PANC-1 cells;however,no significant effects on hTERT-HPNE cells were observed.TQ significantly reduced the mRNA and protein expression levels of HIF-1αin PANC-1,AsPC-1,and BxPC-3 cells.TQ significantly inhibited the expression of the HIF-1αinitial expression pathway(PI3K/AKT/mTOR)related proteins,and promoted the ubiquitination degradation of the HIF-1αprotein in PANC-1 cells.TQ had no effect on the hydroxylation and von Hippel Lindau protein mediated ubiquitination degradation of the HIF-1αprotein but affected the stability of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90,thus promoting its ubiquitination degradation.CONCLUSION The regulatory mechanism of TQ on HIF-1αprotein expression in PC cells was mainly to promote the ubiquitination degradation of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90;Secondly,TQ reduced the initial expression of HIF-1αprotein by inhibiting the PI3K/AKT/mTOR pathway.

Semaphorin 7A impairs barrier function in cultured human corneal epithelial cells in a manner dependent on nuclear factor-kappa B

●AIM:To evaluate the role of semaphorin 7A(Sema7A)and its associated regulatory mechanisms in modulating the barrier function of cultured human corneal epithelial cells(HCEs).●METHODS:Barrier models of HCEs were treated with recombinant human Sema7A at concentrations of 0,125,250,or 500 ng/mL for 24,48,or 72h in vitro.Transepithelial electrical resistance(TEER)as well as Dextran-fluorescein isothiocyanate(FITC)permeability assays were conducted to assess barrier function.To quantify tight junctions(TJs)such as occludin and zonula occludens-1(ZO-1)at the mRNA level,reverse transcriptionpolymerase chain reaction(RT-PCR)analysis was performed.Immunoblotting was used to examine the activity of the nuclear factor-kappa B(NF-κB)signaling pathway and the production of TJs proteins.Immunofluorescence analyses were employed to localize the TJs.Enzyme-linked immunosorbent assay(ELISA)and RT-PCR were utilized to observe changes in interleukin(IL)-1βlevels.To investigate the role of NF-κB signaling activation and IL^(-1)βin Sema7A’s anti-barrier mechanism,we employed 0.1μmol/L IκB kinase 2(IKK2)inhibitor IV or 500 ng/mL IL^(-1)receptor(IL-1R)antagonist.●RESULTS:Treatment with Sema7A resulted in decreased TEER and increased permeability of Dextran-FITC in HCEs through down-regulating mRNA and protein levels of TJs in a time-and dose-dependent manner,as well as altering the localization of TJs.Furthermore,Sema7A stimulated the activation of inhibitor of kappa B alpha(IκBα)and expression of IL-1β.The anti-barrier function of Sema7A was significantly suppressed by treatment with IKK2 inhibitor IV or IL-1R antagonists.●CONCLUSION:Sema7A disrupts barrier function through its influence on NF-κB-mediated expression of TJ proteins,as well as the expression of IL-1β.These findings suggest that Sema7A could be a potential therapeutic target for the diseases in corneal epithelium.

HMGB1基因敲除通过抑制TLR4/NF-κB通路减轻脓毒症小鼠急性肺损伤

目的 研究高迁移率族蛋白B1(HMGB1)基因敲除减轻脓毒症小鼠急性肺损伤及抑制Toll样受体4(TLR4)/核因子-κB(NF-κB)通路的作用。方法 野生型(WT)小鼠分为WT-Sham组和WT-模型组,HMGB1基因敲除(KO)小鼠分为KO-Sham组和KO-模型组。WT-模型组和KO-模型组采用盲肠结扎穿孔术制备脓毒症ALI模型,WT-Sham组和KO-Sham组进行假手术操作。造模后24 h,检测动脉血氧分压(PaO_(2)),计算氧合指数(OI),检测肺组织病理改变,计算肺损伤评分,检测血清及肺组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6、活性氧簇(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)的浓度,肺组织中HMGB1、TLR4、核NF-κB的表达。结果 WT-模型组的PaO_(2)、OI、血清及肺组织SOD的浓度低于WT-Sham组,肺损伤评分、血清及肺组织中TNF-α、IL-1β、IL-6、ROS、MDA的浓度、肺组织中HMGB1、TLR4、核NF-κB的表达水平高于WT-Sham组(P<0.05);KO-模型组肺组织中不表达HMGB1,PaO_(2)、OI、血清及肺组织SOD的浓度高于WT-模型组,肺损伤评分、血清及肺组织中TNF-α、IL-1β、IL-6、ROS、MDA的浓度、肺组织中TLR4、核NF-κB的表达水平低于WT-模型组(P<0.05)。结论敲除HMGB1减轻脓毒症小鼠ALI,相关的分子机制可能是抑制TLR4/NF-κB通路介导的炎症反应和氧化应激反应。

五味子乙素通过TLR4/NF-κB信号通路对急性胰腺炎大鼠肺部损伤的影响

目的:探讨五味子乙素通过Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路对急性胰腺炎(AP)大鼠肺部损伤的影响。方法:取SD大鼠,通过胆胰管内逆行注射5%牛磺胆酸钠方法诱导建立AP肺损伤模型,经随机数表法分为模型组、五味子乙素组、TLR4过表达载体组、TLR4空载组、五味子乙素+TLR4过表达载体组,每组12只大鼠,再取12只SD大鼠仅翻动肠管不注射5%牛磺胆酸钠,作为假手术组。以药物分别干预大鼠后,检测各组大鼠肺功能及各组大鼠腹水量与肺组织湿重/干重(W/D);HE染色检测各组大鼠肺组织病理形态并评分;检测各组大鼠动脉血气;全自动生化分析仪检测大鼠血清淀粉酶,ELISA检测炎症细胞因子IL-6、IL-18水平;蛋白免疫印迹法检测肺组织TLR4/NF-κB通路蛋白表达;免疫组织化学染色检测肺组织TLR4蛋白表达。结果:与假手术组相比,模型组大鼠肺组织出现病理损伤改变,模型组大鼠MV、PEF、PaO_(2)、OI显著降低(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平显著升高(P<0.05)。与模型组、五味子乙素+TLR4过表达载体组分别相比,五味子乙素组大鼠肺组织病理损伤改变程度均减轻,MV、PEF、PaO_(2)、OI均升高(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平均降低(P<0.05);TLR4过表达载体组大鼠肺组织病理损伤改变程度均加重,MV、PEF、PaO_(2)、OI均降低(P<0.05),Ri、腹水量与W/D、PaCO_(2)、Holfbauer评分、血清淀粉酶、IL-6与IL-18水平、肺组织TLR4阳性细胞比例、TLR4与MYD88蛋白表达、p-NF-κB p65/NF-κB p65水平均升高(P<0.05)。与模型组相比,TLR4空载组大鼠各指标差异无统计学意义(P>0.05)。结论:五味子乙素可通过下调TLR4/NF-κB信号通路,抑制炎症,减轻AP大鼠肺部损伤,修复肺功能。

左归降糖舒心方调控TLR4/NF-κB通路抑制糖尿病心肌病小鼠心肌纤维化的机制研究

目的 基于Toll样受体4(Toll-like receptor 4,TLR4)/核因子κB(nuclear factor-κB,NF-κB)信号通路探讨左归降糖舒心方对糖尿病心肌病MKR小鼠心肌纤维化及炎症因子的影响。方法 以8周龄雄性MKR小鼠为实验对象,采用高脂饮食联合腹腔注射链脲佐菌素(streptozotocin,STZ)40 mg/kg构建糖尿病心肌病模型,随机分为中药高剂量组[33.67 g/(kg·d)左归降糖舒心方2 g/mL]、中药低剂量组[16.84 g/(kg·d)左归降糖舒心方2 g/mL]、西药联合组[0.23 g/(kg·d)二甲双胍联合1.5 mg/(kg·d)依那普利]和模型组(等容量蒸馏水);另设FVB小鼠为空白对照组(等容量蒸馏水)。每组10只,连续给药8周后取材。收集尾静脉血液检测空腹血糖水平;采用HE、Masson染色观察心肌病理改变,电镜观察心肌组织超微结构变化;采用ELISA检测小鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及白细胞介素-1β(interleukin-1β,IL-1β)含量;采用Western blot法检测TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达情况;RT-qPCR及Western blot检测小鼠心肌组织Ⅰ型胶原(collagen type I,CollagenⅠ)、Ⅲ型胶原(collagen typeⅢ,CollagenⅢ)及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达水平。结果 与空白对照组比较,模型组小鼠空腹血糖及血清中TNF-α、IL-1β含量增高(P<0.01);心肌细胞结构紊乱,胶原含量增加,心肌细胞重度退行性变;心肌组织中TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达上调(P<0.01),CollagenⅠ、CollagenⅢ、α-SMA mRNA及蛋白表达增加(P<0.05,P<0.01),中药高剂量组小鼠心肌组织CollagenⅠ、CollagenⅢ蛋白表达增加(P<0.01)。与模型组比较,中药高、低剂量组和西药联合组小鼠空腹血糖及血清中TNF-α、IL-1β含量均降低(P<0.01);心肌结构改善,胶原沉积减少;心肌组织中TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达下调(P<0.01),CollagenⅠ、CollagenⅢ、α-SMA蛋白及mRNA表达下调(P<0.01)。与西药联合组比较,中药低剂量组小鼠空腹血糖及血清中TNF-α、IL-1β含量均升高(P<0.01),心肌结构无明显改善,胶原沉积无显著减少;心肌组织中TLR4、NF-κB p56、p-NF-κB p56/NF-κB p56蛋白表达上调(P<0.01),CollagenⅠ、CollagenⅢ、α-SMA蛋白及mRNA表达上调(P<0.01)。结论 左归降糖舒心方能抑制心肌炎性反应、改善心肌细胞结构,其作用机制可能与调控TLR4/NF-κB通路、下调细胞炎症因子表达、抑制心肌纤维化有关。

姜黄素抑制NF-κB信号通路缓解氧化应激对成骨分化的损害发挥抗骨质疏松作用

目的 探讨姜黄素抑制氧化应激对成骨分化损害的机制及以剂量依赖的方式发挥抗骨质疏松的作用。方法 采用细胞氧化应激模型,加入不同浓度的姜黄素,测定骨形成指标,并检测参与的潜在信号通路。同时,用姜黄素处理小鼠去卵巢(ovariectomized, OVX)骨质疏松动物模型来证实其抗骨质疏松的作用。结果 体外实验发现,低浓度姜黄素(1~10μmol·L^(-1))促进成骨细胞增殖,提高骨形成碱性磷酸酶(alkaline phosphatase, ALP)活性,逆转氧化应激导致的成骨钙沉积下降,降低了核因子kappa-B配体的受体激动剂(RANKL)和白介素-6 (IL-6)的表达。体内实验结果显示,姜黄素(5 mg·kg^(-1))给药后部分逆转了OVX小鼠血液中丙二醛(malondialdehyde, MDA)和谷胱甘肽(glutathione, GSH)活性的比例、降低高骨代谢、增加骨密度(bone density, BMD)、改善了骨小梁的微结构,但高浓度姜黄素无氧化应激保护作用。结论 姜黄素可以减轻氧化应激的骨形成损害,NF-κB信号通路是主要参与通路,姜黄素可能是预防骨质疏松症的理想药物。

白细胞介素6与核因子κB对胰腺癌的诊断价值及其与临床病理分期的关系研究

目的探究白细胞介素6(IL-6)、核因子κB(NF-κB)对胰腺癌的诊断价值及其与胰腺癌临床病理分期的关系。方法收集2019年6月至2022年6月新疆维吾尔自治区人民医院收治的68例胰腺癌患者作为病例组,60例同期于本院门诊健康体检者作为对照组。检测2组受试者血清中IL-6、NF-κB的表达水平,采用受试者工作特征(ROC)曲线分析IL-6、NF-κB表达对胰腺癌诊断的价值;比较不同临床病理分期患者血清IL-6、NF-κB水平的表达差异。结果病例组患者血清IL-6和NF-κB水平明显高于对照组,差异有统计学意义[IL-6:16.33(8.41,67.41)ng/L比6.15(5.55,7.33)ng/L,Z=6.109,P<0.001;NF-κB:449.52(230.37,568.28)ng/L比150.35(126.48,167.30)ng/L,Z=5.463,P<0.001]。IL-6、NF-κB诊断胰腺癌ROC曲线下面积(AUC)分别为0.813±0.041、0.920±0.024,联合检测的AUC为0.923±0.024。T_(3)、T_(4)期胰腺癌患者血清IL-6、NF-κB水平与T_(1)、T_(2)期患者比较差异均有统计学意义(均P<0.05)。血清IL-6、NF-κB水平随着淋巴结转移程度(N_(0)~N_(2))的增高而增高,差异均有统计学意义(均P<0.05),且N_(1)、N_(2)期胰腺癌患者血清IL-6、NF-κB水平与N_(0)期患者相比,差异均有统计学意义(均P<0.05)。远处转移胰腺癌患者(M_(1))的血清IL-6、NF-κB水平高于未远处转移的患者(M_(0)),差异有统计学意义(P=0.024)。结论血清IL-6、NF-κB可以作为胰腺癌早期诊断、临床病理分期的血清学标志物,其可能与胰腺癌的发生、发展密切相关。

黄芪阳和汤调控PI3K/AKT/NF-κB信号通路促进糖尿病足溃疡大鼠创面愈合

目的基于磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/核因子-κB(NF-κB)信号通路探究黄芪阳和汤对糖尿病足溃疡(DFU)大鼠创面愈合的影响。方法构建DFU大鼠模型,将建模成功的48只大鼠随机分为模型组,黄芪阳和汤低(8.5 g/kg)、高(17 g/kg)剂量组,黄芪阳和汤高剂量(17 g/kg)+LY294002(PI3K/AKT通路抑制剂,0.3 mg/kg)组;每组12只;另取12只大鼠为对照组。各组大鼠给予对应药物干预,连续4周。第14、28天给药后,观察大鼠一般状态及创面变化,计算创面愈合率,检测大鼠空腹血糖(FBG)水平和大鼠创面周围组织经皮氧分压(TcpO2);酶联免疫吸附试验检测大鼠血清血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)、C反应蛋白(CRP)、白细胞介素(IL)-6水平;苏木素-伊红染色观察大鼠创面组织病理学变化;免疫组织化学染色测定大鼠创面组织微血管密度;蛋白免疫印迹法检测大鼠创面组织中PI3K、磷酸化PI3K(p-PI3K)、AKT、磷酸化AKT(p-AKT)、NF-κB p65、磷酸化NF-κB p65(p-NF-κB p65)、NF-κB抑制蛋白α(IκB-α)蛋白表达。结果对照组大鼠毛色光滑,饮食、饮水、排泄均正常,较活跃,创面愈合快,创面组织炎症反应较轻,新生血管较多,肉芽组织中成纤维细胞及胶原基质丰富;模型组大鼠毛色暗淡无光泽,活动减少,且出现多饮、多食、多尿症状,创面颜色较深,且周围组织出现水肿、溃疡,创面组织可见大量炎性细胞浸润,伴组织坏死、渗出,新生血管及成纤维细胞较少,创面愈合率、创面周围组织TcpO2、血清VEGF、HIF-1α、创面组织微血管密度、p-PI3K、p-AKT、IκB-α蛋白表达水平降低,FBG、血清CRP、IL-6、创面组织p-NF-κB p65蛋白表达升高(P<0.05);与模型组相比,黄芪阳和汤低、高剂量组大鼠状态逐渐改善,创面组织病变程度依次减轻,创面愈合率、创面周围组织TcpO2、血清VEGF、HIF-1α、创面组织微血管密度、p-PI3K、p-AKT、IκB-α蛋白表达水平依次升高,FBG、血清CRP、IL-6、创面组织p-NF-κB p65蛋白表达依次降低(P<0.05);LY294002能部分逆转高剂量黄芪阳和汤对DFU大鼠的治疗作用(P<0.05)。结论黄芪阳和汤能调控PI3K/AKT/NF-κB信号通路,抑制DFU大鼠炎症反应,促进血管新生,从而促进创面愈合。

淫羊藿苷调控mTOR/Akt/CREB通路对高糖诱导的足细胞自噬及凋亡的影响

目的 探讨淫羊藿苷对高糖诱导的足细胞自噬、凋亡及哺乳动物雷帕霉素靶蛋白(mTOR)/丝氨酸苏氨酸蛋白激酶(Akt)/环磷酸腺苷反应元件结合蛋白(CREB)通路的影响。方法 将小鼠足细胞MPC5分为5组:正常对照组(5.5 mmol·L^(-1)葡萄糖)、高糖组(30 mmol·L^(-1)葡萄糖)、淫羊藿苷组(30 mmol·L^(-1)葡萄糖+5μmol·L^(-1)淫羊藿苷)、GDC-0349组(30 mmol·L^(-1)葡萄糖+50μmol·L^(-1)GDC-0349)、淫羊藿苷+GDC-0349组(30 mmol·L^(-1)葡萄糖+5μmol·L^(-1)淫羊藿苷+50μmol·L^(-1)GDC-0349)。培养48 h后,噻唑蓝法检测MPC5细胞活力;吖啶橙染色观察MPC5细胞自噬情况;流式细胞术检测MPC5细胞凋亡;蛋白印迹法检测MPC5细胞自噬[微管相关蛋白1轻链3(LC3)Ⅱ、LC3Ⅰ、自噬相关蛋白(Beclin-1)]、凋亡[Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)]和mTOR/Akt/CREB通路相关蛋白的表达。结果 与正常对照组比较,高糖组MPC5细胞活力、Bcl-2、磷酸化mTOR(p-mTOR)/mTOR、磷酸化Akt(p-Akt)/Akt、磷酸化CREB(p-CREB)/CREB蛋白表达水平显著降低(P<0.05),自噬能力增强,自噬体表现出橙色荧光,细胞凋亡率、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bax蛋白表达水平显著升高(P<0.05)。与高糖组比较,淫羊藿苷组MPC5细胞活力、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bcl-2、p-mTOR/mTOR、p-Akt/Akt、p-CREB/CREB蛋白表达水平显著升高,自噬能力进一步增强,自噬体数量增多,自噬体呈现出砖红色荧光(P<0.05),细胞凋亡率、Bax蛋白表达水平显著降低(P<0.05);GDC-0349组MPC5细胞活力、LC3Ⅱ/LC3Ⅰ、Beclin-1、Bcl-2、p-mTOR/mTOR、p-Akt/Akt、p-CREB/CREB蛋白表达水平显著降低,自噬能力减弱,自噬体数量减少,自噬体表现出橙色荧光(P<0.05),细胞凋亡率、Bax蛋白表达水平显著升高(P<0.05);淫羊藿苷+GDC-0349可逆转淫羊藿苷对高糖诱导MPC5细胞的作用效果(P<0.05)。结论 淫羊藿苷通过激活mTOR/Akt/CREB通路促进高糖诱导的足细胞自噬抑制细胞凋亡。

基于线粒体Cyt b基因序列的4个黄尾鲴养殖群体遗传多样性分析

黄尾鲴(Xenocypris davidi)是浙江省自然水域增殖放流的主要鱼类,为了解人工繁育对黄尾鲴遗传多样性的影响,利用线粒体DNA细胞色素b基因(Cyt b)对4个养殖群体的遗传多样性进行研究,旨在为黄尾鲴增殖放流策略制定和实施提供基础数据。结果显示,在编码Cyt b的1 140 bp序列中,检测到157个变异位点,界定了21种单倍型,其中长兴、双浦、八里店和醴陵群体的单倍型数目分别为10个、11个、7个和2个,单倍型多样性介于0.226~0.794,核苷酸多样性介于0.006 14~0.023 86。除醴陵群体遗传多样性较低外,其余3个养殖群体的遗传多样性具有高单倍型数和高核苷酸多样性的特点。4个黄尾鲴养殖群体间的遗传距离为0.018 74~0.092 74,遗传分化指数为0.808 63(P<0.01),其中长兴和八里店群体的分化程度较低,双浦和醴陵群体的分化程度较高,且遗传变异主要发生在群体间。本研究结果可从分子水平为黄尾鲴的资源保护和人工增殖放流提供参考依据。

以皮肤肿块为首发临床表现的伴MYC和BCL2重排弥漫性大B细胞淋巴瘤

报告1例以皮肤肿块为首发临床表现的伴细胞性骨髓细胞瘤病病毒癌(MYC)基因和B细胞淋巴瘤因子2(BCL2)基因重排弥漫性大B细胞淋巴瘤。患者女,87岁。左侧眉弓上方皮肤结节1年。皮肤科检查:左眉上方一肤色圆形肿物,隆起于皮肤表面,肿物表面可见浸润性红斑,伴少许渗出,并见结痂、鳞屑,边界清楚。皮损组织病理检查:表皮形态正常,真皮层内大量母细胞样淋巴细胞结节状浸润。免疫组化:母细胞样淋巴细胞CD45、CD79a、B细胞淋巴瘤因子6(BCL6)、细胞周期蛋白D1(cyclin D1)及MYC均(+);CD3、CD2、CD30、细胞角蛋白(CKP)、间变性淋巴瘤激酶(ALK)及Epstein-Barr病毒编码RNA(EBER)均(-)。荧光原位杂交检测:MYC、BCL2及BCL6均重排。诊断:伴MYC和BCL2重排弥漫性大B细胞淋巴瘤。患者未治疗,2个月后死亡。

血友病B治疗药物情报分析

血友病B(HB)是一种X染色体隐性遗传出血性疾病。过去几十年来,HB治疗取得了巨大进展。从凝血酶原复合物到血源性凝血因子提高了治疗安全性,从标准半衰期重组凝血因子到生物工程长效重组凝血因子改善了患者依从性和临床结局。非因子替代疗法不但能够通过皮下给药为患者减轻治疗负担,还避免了产生抑制物的风险,同时为已产生抑制物的患者提供了新的治疗方案。腺相关病毒载体基因治疗开启了HB的新篇章,目前已有一种基因治疗在美国获批上市。该文通过对欧美等发达国家及中国上市药物信息追踪,对在研药物研发管线及主要研究成果等信息进行挖掘,对HB治疗药物进行全视角、全周期情报解读,以期为医疗机构及相关研发单位提供参考。

B群链球菌暴露后蜕膜间质细胞死亡和炎症信号的相关性研究

目的探究B群链球菌暴露后蜕膜间质细胞死亡和炎症信号的相关性。方法选取2017年10月至2019年10月于保定市第二中心医院妇产科分娩的孕妇100例,分为早产组27例和足月顺产组73例,取孕妇阴道拭子和蜕膜间质组织,检测B群链球菌阳性率、蜕膜间质细胞凋亡情况、血清炎性因子水平、NLRP3、Caspase-1、OX40、OX40L、PD-1和PD-L1 mRNA表达,分析B群链球菌暴露后蜕膜间质细胞死亡和炎症信号的相关性。结果2组孕妇B群链球菌阳性率、蜕膜间质细胞凋亡情况、血清炎性因子水平、NLRP3、Caspase-1、OX40、OX40L、PD-1和PD-L1 mRNA表达比较,差异均有统计学意义(P<0.05)。早产组孕妇的GBS阳性率和蜕膜组织细胞凋亡值分别为37.04%和(46.73±4.92),足月顺产组的GBS阳性率和蜕膜组织细胞凋亡值仅为9.59%和(6.03±2.04);早产组孕妇的血清炎性因子水平高于足月顺产组,蜕膜间质细胞凋亡的孕妇血清炎性因子水平高于未凋亡的孕妇,差异有统计学意义(P<0.05)。结论B群链球菌暴露后蜕膜间质细胞死亡和炎症信号存在相关性,蜕膜间质细胞死亡会引发炎性因子水平升高。

罕见抗-Di^(b)致严重胎儿新生儿溶血病的实验室检测与相关研究

目的对1例高频抗体导致的胎儿新生儿溶血病(hemolytic disease of the fetus and newborn,HDFN)进行检测、鉴定及配血。方法对患儿进行新生儿溶血试验,对母亲进行血清学意外抗体鉴定,并对母亲红细胞进行常见高频抗原鉴定;对检出抗体进行IgG分型检测,并用流式细胞术进行单核细胞体外吞噬致敏红细胞试验,以检测抗体相关的吞噬率;对患儿母亲、父亲及舅舅进行相关红细胞血型基因测序;利用稀释的母亲血浆和抗人球卡法,在献血者中进行大规模相合血液的筛选。结果产妇鉴定为Di(b-)稀有血型,产生了抗-Di b(效价512)并导致了严重的HDFN;抗-Di b亚型分型为IgG1和IgG2型,单核细胞体外吞噬效率为88.83%(74.7/84.09);产妇亲属中没有相合献血者,后续从5520名献血者中筛选到2例Di(b-)相合血液,患儿接收输血治疗后康复出院。后续在51334名献血者中筛查到17名Di(b-)献血者,该数据表明Di(b-)在广州地区献血者中的分布频率约为三千分之一(0.033%,17/51334)。结论综合利用血型血清学及分子生物学方法诊断了抗-Di b所致的严重HDFN,建立了1种有效大规模筛查Di(b-)稀有血型的方法并找到相合血液,为建立Di(b-)稀有血型库奠定了基础。

分数[a,b]-因子的紧孤立韧度条件

本文研究了分数[a,b]-因子和孤立韧度相关性的问题.利用子图分解的方法,获得了一个图存在分数[a,b]-因子的孤立韧度条件,通过反例说明该条件是紧的.改进了原有对分数[a,b]-因子的孤立韧度界.

钢渣/氮掺杂改性活性炭催化过硫酸盐降解罗丹明B

以提高钢渣资源化利用和深度降解有机污染物为研究目标,通过改变氨气煅烧温度制备了系列钢渣/氮掺杂改性活性炭复合材料,并用于催化过硫酸钠处理罗丹明B(RhB)废水。同时采用N2吸附/脱附、XRD、XPS等手段对复合材料的孔结构和表面性质进行了表征。结果表明:氮元素的引入可改善复合材料中活性金属与活性炭之间的相互作用,从而提高复合材料的催化活性。以最优复合材料60%GZ/AC-N800为研究对象,在反应温度为35℃、PS加量为2 g/L和宽pH(3~11)操作条件下,反应30 min后罗丹明B(100 mg/L)的去除率可达88.7%~92.2%。动力学分析发现,60%GZ/AC-N800复合材料催化PS降解RhB符合准二级动力学。自由基掩蔽实验发现,该降解过程是自由基途径(SO_(4)^(·-)和·OH)和非自由基途径(^(1)O_(2))共同参与反应的氧化过程。

B亚型新等位基因803delC的分子生物学研究

目的分析研究1例新的B亚型等位基因血清学特点和分子生物学机制。方法应用血清学方法检测患者ABO血型。应用序列特异性引物-聚合酶链反应(PCR-SSP)检测ABO血型基因。应用Sanger基因序列分析检测ABO基因1~7外显子编码区域,确定基因突变位点。结果血清学鉴定患者正定型为O型,反定型为B型。PCR-SSP基因分型结果为A/O型,存在A基因,与血清学结果不符。进一步Sanger双链测序结果显示该标本在ABO^(*)B.01/ABO^(*)O.01.01的基础上,第7外显子803位置缺失C碱基。该突变最终导致多肽链上发生p.Ala268Gly和p.Phe269Ser的氨基酸替换,并且从269位置开始产生新的开放阅读框,新的开放阅读框第20号氨基酸为终止密码子,导致B基因表达终止。进一步ABO基因克隆测序证明该突变点位于ABO*B.01基因上,该突变已提交NCBI数据库,收录编号为OR343908。结论在中国人群中发现1种新的导致B变异型的ABO等位基因,基因检测方法可辅助鉴定血清学正、反定型不符的疑难血型。

多黏菌素B治疗危重症儿童多重耐药革兰阴性菌感染的疗效观察

目的:评估多黏菌素B治疗危重症儿童多重耐药革兰阴性菌感染的疗效,并探讨其影响多黏菌素B疗效的可能因素。方法:选取某院重症监护室2020年2月~2022年6月诊治的26例危重症多重耐药革兰阴性菌感染患儿,根据患者的疗效分为临床有效组(n=14)和临床无效组(n=12)。比较两组患者的性别、年龄、小儿危重病例评分、合并疾病、病原菌、治疗时间、机械通气情况以及与其他抗菌药物联用情况。对其中有意义的变量进行多因素Logistic回归分析,探讨影响多黏菌素B疗效的可能因素。结果:临床有效组与无效组的性别、年龄、小儿危重病例评分、合并疾病、病原菌以及与其他抗菌药物联用情况比较均无统计学差异(P>0.05)。临床有效组的多黏菌素B使用时间较无效组更长(P=0.000),临床无效组有更多的患儿应用机械通气(P=0.034)。Logistic回归分析果结提示治疗时间(OR:2.606,95%CI:1.346~5.046,P=0.004)是影响多黏菌素B治疗危重症儿童多重耐药革兰阴性菌感染临床疗效的可能因素。结论:多黏菌素B治疗危重症儿童多重耐药革兰阴性菌感染具有较好的治疗效果,治疗时间是影响多黏菌素临床疗效的可能因素。