目的 探讨骨碎补(rhizoma drynariae,Rhd)对骨髓腔内脂肪异常积累引起的脂毒性介导成骨细胞焦亡的影响及其机制.方法 体内构建卵巢摘除(ovariectomy,OVX)骨质疏松小鼠模型,用Rhd进行灌胃,给药 8 周后称重,取股骨组织和血清.HE染色观察髓腔内脂肪堆积状态.体外构建成脂-成骨细胞共培养系统,茜素红S(alizarin red S,ARS)染色观察钙化结节的数量,碱性磷酸酶(alkaline phosphatease,ALP)染色检测碱性磷酸酶活性水平,乳酸脱氢酶(lactate dehydrogenase,LDH)检测和Hoechst33342/PI染色检测共培养下成骨细胞(osteoblasts,OBs)焦亡水平,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞培养上清液中白细胞介素-1β(interleukin-1β,IL-1β)和白细胞介素-18(interleukin-18,IL-18)的释放量,Western blot检测成骨能力相关蛋白Runt相关转录因子 2(runt-related transcription factor 2,RUNX2)、骨形态发生蛋白 2(bone morphogenetic protein,BMP2)和 1 型胶原蛋白(collagen-1,COL-1).选择NLRP3 特异性抑制剂MCC950 作为阳性对照检测焦亡相关蛋白NOD样受体热蛋白结构域相关蛋白 3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、半胱天冬酶-1(cysteinyl aspartate specific proteinase-1,CASP-1)、效应蛋白消皮素D(gasdermin D,GSDMD)、IL-1β和IL-18 的表达.结果 OVX小鼠髓腔内发生骨质流失伴大量脂肪堆积,Rhd灌胃可以有效缓解这一趋势.在成脂-成骨共培养系统中,共培养环境抑制了OBs活性和OBs中ALP 活性及矿化结节,抑制了RUNX2、BMP2、COL-1 蛋白表达,并且诱导焦亡发生,提高了LDH释放量和PI染色细胞阳性率,以及上调NLRP3、CASP-1、GSDMD、IL-1β和IL-18 蛋白表达水平.而这一趋势在Rhd干预后得到逆转.结论 大量脂肪堆积引起的脂毒性可以抑制OBs活性并通过激活NLRP3 炎症小体诱导OBs焦亡,而Rhd可能通过抑制NLRP3 炎症小体的激活以抑制OBs焦亡和炎性反应,从而起到抗骨质疏松的作用.

Objective:Icariin(ICA)has a good neuroprotective effect and can upregulate neuronal basal autophagy in naturally aging rats.Mitochondrial dysfunction is associated with brain aging-related neurodegenerative diseases.Abnormal opening of the mitochondrial permeability transition pore(mPTP)is a crucial factor in mitochondrial dysfunction and is associated with excessive autophagy.This study aimed to explore that ICA protects against neuronal injury by blocking the mPTP opening and down-regulating autophagy levels in a D-galactose(D-gal)-induced cell injury model.Methods:A cell model of neuronal injury was established in rat pheochromocytoma cells(PC 12 cells)treated with 200 mmol/L D-gal for 48 h.In this cell model,PC12 cells were pre-treated with different concentrations of ICA for 24 h.MTT was used to detect cell viability.Senescence associated β-galactosidase(SA-β-Gal)staining was used to observe cell senescence.Western blot analysis was performed to detect the expression levels of a senescence-related protein(p21),autophagy markers(LC3B,p62,Atg7,Atg5 and Beclin 1),mitochondrial fission and fusion-related proteins(Drp1,Mfn2 and Opa1),and mitophagy markers(Pink1 and Parkin).The changes of autophagic flow were detected by using mRFP-GFP-LC3 adenovirus.The intracellular ultrastructure was observed by transmission electron microscopy.Immunofluorescence was used to detect mPTP,mitochondrial membrane potential(MMP),mitochondrial reactive oxygen species(mtROS)and ROS levels.ROS and apoptosis levels were detected by flow cytometry.Results:D-gal treatment significantly decreased the viability of PC12 cells,and markedly increased the SA-β-Gal positive cells as compared to the control group.With the D-gal stimulation,the expression of p21 was significantly up-regulated.Furthermore,D-gal stimulation resulted in an elevated LC3B Ⅱ/Ⅰ ratio and decreased p62 expression.Meanwhile,autophagosomes and autolysosomes were significantly increased,indicating abnormal activation of autophagy levels.In addition,in this D-gal-induced model of cell injury,the mPTP was abnormally open,the ROS generation was continuously increased,the MMP was gradually decreased,and the apoptosis was increased.ICA effectively improved mitochondrial dysfunction to protect against D-gal-induced cell injury and apoptosis.It strongly inhibited excessive autophagy by blocking the opening of the mPTP.Cotreatment with ICA and an mPTP inhibitor(cyclosporin A)did not ameliorate mitochondrial dysfunction.However,the protective effects were attenuated by cotreatment with ICA and an mPTP activator(lonidamine).Conclusion:ICA inhibits the activation of excessive autophagy and thus improves mitochondrial dysfunction by blocking the mPTP opening.

目的:探讨青风藤活性成分对糖尿病(diabetes mellitus,DM)db/db小鼠肾脏的保护作用及其机制研究.方法:30 只 6 周龄雄性db/db小鼠随机分为模型组(等体积生理盐水),阳性组(195 mg/kg/d羟苯磺酸钙),盐酸青藤碱(Sinomenine Hydrochloride,SH)低、中、高剂量组(31.2、62.4、124.8 mg/kg/d SH)5 组,同时选取 6 只同周龄db/m小鼠设为对照组(等体积生理盐水),每日 1 次,定时灌胃给药,连续 6w.于 12w末观察各组小鼠一般情况,检测尿微量白蛋白(mAlb)、血肌酐(SCr)和血尿素氮(BUN)水平;试剂盒检测肾组织匀浆中丙二醛(MDA)、还原型谷胱甘肽(GSH)和超氧化物歧化酶(SOD)的水平;Luminex多因子检测技术测定血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和白细胞介素-10(IL-10)的含量;ELISA法检测血清免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、免疫球蛋白A(IgA)及补体蛋白3(C3)的水平;苏木精-伊红(HE)和过碘酸-雪夫(PAS)染色观察肾脏组织病理形态变化;透射电镜观察肾脏超微结构的改变.结果:与模型组比较,阳性组和SH低、中、高剂量组小鼠mAlb、SCr和BUN水平均降低(P＜0.05);肾组织匀浆中MDA水平降低(P＜0.05),GSH水平和SOD活性均升高(P＜0.05);血清中TNF-α、IL-1β、IL-6、IgG、IgM、IgA和补体C3 水平均降低(P＜0.05),IL-10 水平升高(P＜0.05);镜下肾小球损伤及肾小管上皮水肿情况均有所改善,肾组织损伤减轻.结论:青风藤活性成分对DM db/db小鼠的肾脏具有保护作用,其作用机制可能与改善氧化应激反应,缓解炎症反应及提高免疫功能有关.

成骨细胞失调所导致的骨形成机制障碍是临床上常见的骨代谢相关疾患的病理基础.研究表明,Wnt/β-连环蛋白(Wnt/β-catenin)、磷脂酰基醇-3-激酶/蛋白激酶B(PI3K/AKT)、核因子E2 相关因子 2/血红素加氧酶-1(Nrf2/HO-1)、腺苷酸活化蛋白激酶(AMPK)、沉默信息调节因子 6/核转录因子-κB(SIRT6/NF-κB)等信号通路均具有调节成骨细胞,维持骨稳态的能力.而二甲双胍作为临床一线降糖药物不仅具有降糖能力,而且在调控成骨细胞增殖分化等过程中发挥重要作用.笔者通过总结上述信号通路与二甲双胍干预的研究现状,旨在为成骨细胞相关研究提供新思路,为改善骨形成促进骨稳态提供理论参考与依据.

Objective:Mitofusin-2(MFN2)is a mitochondrial membrane protein that plays a critical role in regulating mitochondrial fusion and cellular metabolism.To further elucidate the impact of MFN2,this study aimed to investigate its significance on hepatocellular carcinoma(HCC)cell function and its potential role in mediating chemosensitivity.Methods:This study investigated the effects of silencing and overexpressing MFN2 on the survival,proliferation,invasion and migration abilities,and sorafenib resistance of MHCC97-L HCC cells.Additional experiments were conducted using XAV939(a β-catenin inhibitor)and HLY78(a β-catenin activator)to further validate these findings.Results:Silencing MFN2 significantly promoted the survival and proliferation of MHCC97-L cells,enhanced their invasion and migration capacities,increased the IC50 of sorafenib,reduced the percentage of TUNEL-positive cells,and decreased the expression of proapoptotic proteins.Additionally,silencing MFN2 markedly induced the nuclear translocation of β-catenin,increased β-catenin acetylation levels and enhanced the expression of the downstream regulatory proteins Snail 1 and Vimentin while inhibiting E-cadherin expression.Conversely,overexpressing MFN2 reversed the effects observed in MHCC97-L cells mentioned above.The results confirmed that silencing MFN2 activated the β-catenin/epithelial-mesenchymal transition(EMT)pathway and reduced the sensitivity of cells to sorafenib,which could be reversed by XAV939 treatment.Conversely,overexpression of MFN2 inhibited the β-catenin/EMT pathway and increased the sensitivity of cells to sorafenib,which could be altered by HLY78.Conclusion:Low expression of MFN2 in HCC cells promotes the nuclear translocation of β-catenin,thereby activating the EMT pathway and mediating resistance to sorafenib.

目的 观察骨质疏松性椎体压缩性骨折(osteoporotic vertebral compression fracture,OVCF)患者在经皮椎体成形术后使用特立帕肽序贯地舒单抗对比序贯唑来膦酸治疗绝经后骨质疏松症的疗效.方法 回顾性选择 2020 年 12 月至 2023 年 12月在南京市第一医院行经皮椎体成形术且术后使用特立帕肽治疗 12 个月的绝经后骨质疏松症患者 60 例,根据序贯用药情况将纳入患者分为特立帕肽序贯地舒单抗组和特立帕肽序贯唑来膦酸组,每组 30 例.比较两组治疗前、治疗 6 个月、治疗 12 个月的血清P1NP、血清β-CTX、腰椎及髋部骨密度(bone mineral density,BMD)变化情况和再骨折的发生情况,评估两组的疗效差异.结果 地舒单抗组治疗 12 个月的髋部BMD增长率高于唑来膦酸组(P=0.001).两组治疗 12 个月后的腰椎和髋部BMD均高于治疗前(P＜0.05),地舒单抗组和唑来膦酸组治疗 12 个月的腰椎BMD较治疗前分别提升 8.7%和 6.7%,髋部BMD较治疗前分别提升 6.6%和 2.9%.地舒单抗组治疗 6 个月、12 个月的血清P1NP和血清β-CTX水平均低于唑来膦酸组(P＜0.05).两组治疗期间再骨折发生率无统计学差异(P＞0.05).结论 合并OVCF的绝经后骨质疏松症患者,特立帕肽序贯地舒单抗较序贯唑来膦酸对提高髋部骨密度和降低骨代谢标志物的作用更明显,两种治疗方案对再骨折的发生率无明显影响.

目的 评估第1秒用力呼气容积(FEV1)、肺一氧化碳弥散量(DLCO)对非小细胞肺癌(NSCLC)患者经胸腔镜肺叶切除术后肺部并发症(PPC)及短期预后的预测价值.方法 前瞻性对 2019 年 2 月—2023 年 1 月在保定市第二中心医院行电视胸腔镜手术肺叶切除术的 150 例NSCLC患者进行分析,根据术后 30d内的PPC发生情况,将患者分为PPC组(n=26)及无PPC组(n=124),并记录患者的院内死亡情况.结果 PPC组患者FEV1%术后预测值(ppoFEV1%)、DLCO%术后预测值(ppoDLCO%)和DLCO更低,而住院时间和院内病死率,吸烟史比例更高,年龄更大(P＜0.05).通过受试者(ROC)曲线分析显示,ppoFEV1%和ppoDLCO%预测肺叶切除术患者发生PPC的曲线下面积(AUC)分别为 0.850(95%CI:0.771～0.930)、0.858(95%CI:0.779～0.936),最佳截断值分别为 59.26%、53.25%.Lasso回归和逐步回归分析显示,年龄(β=0.074,OR=1.077),ppoFEV1%(β=-0.036,OR=0.964)和 ppoDLCO%(β=-0.103,OR=0.902)是具有统计学意义的预测因素(P<0.05).通过Pearson相关性分析,PPC组患者的ppoFEV1%、ppoDLCO%与住院时间呈负相关(r=-0.473,P=0.015;r=-0.434,P=0.027).经ROC曲线分析,ppoFEV1%、ppoDLCO%对肺叶切除术患者院内死亡的预测AUC分别为 0.850(95%CI:0.771～0.930)、0.926(95%CI:0.871～0.982).结论 ppoFEV1%＜59.26%、ppoDLCO%＜53.25%的行电视胸腔镜手术肺叶切除术的NSCLC患者具有更高的PPC发生风险,检测患者的ppoFEV1%、ppoDLCO%有助于临床医生对NSCLC患者进行风险分层、预后预测.

Objective:This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside Ⅳ on key targets using high-throughput sequence technology and bioinformatics analyses.Methods:PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25,0.5,0.75,1,and 1.25 mg/mL for 24 h.Cell morphology was evaluated,and cell survival rates were calculated.A neurocyte inflammatory model was established with LPS treatment,which reached a 50％cell survival rate.PC12 cells were treated with 0.01,0.1,1,10,or 100 μmol/L astragaloside Ⅳ for 24 h.The concentration of astragaloside Ⅳ that did not affect the cell survival rate was selected as the treatment group for subsequent experiments.NOS activity was detected by colorimetry;the expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting.The differentially expressed genes(DEGs)between the groups were screened using a second-generation sequence(fold change＞2,P＜0.05)with the following KEGG enrichment analysis,RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells.Results:The viability of PC12 cells was not altered by treatment with 0.01,0.1,or 1 μmol/L astragaloside Ⅳ for 24 h(P＞0.05).However,after treatment with 0.5,0.75,1,or 1.25 mg/mL LPS for 24 h,the viability steadily decreased(P＜0.01).The mRNA and protein expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS,and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h(P＜0.01);however,these changes were reversed when PC12 cells were pretreated with 0.01,0.1,or 1 μmol/L astragaloside Ⅳ in PC12 cells and then treated with 1 mg/mL LPS for 24 h(P＜0.05).Second-generation sequencing revealed that 1026 genes were upregulated,while 1287 genes were downregulated.The DEGs were associated with autophagy,TNF-α,interleukin-17,MAPK,P53,Toll-like receptor,and NOD-like receptor signaling pathways.Furthermore,PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2,CCL11,CCL7,MMP3,and MMP10,which arc associated with the IL-17 pathway.RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results.Conclusion:LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage.astragaloside Ⅳ plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.

目的:探讨初诊2型糖尿病（T2DM）患者内脏脂肪（VAT）含量与心功能的关系。方法:收集2015年8月至2016年6月就诊的18~70岁初诊T2DM患者，按体质指数（BMI）分为肥胖组（BMI≥28.0 kg/m 2）、超重组（24.0 kg/m 2≤BMI<28.0 kg/m 2）及正常体重组（18.5 kg/m 2≤BMI<24.0 kg/m 2）。采用双能X线骨密度仪测量VAT质量、面积，采用心脏彩超测量右心室前壁脂肪、心尖脂肪及心功能[射血分数、舒张末期容积（EDV）、收缩末期容积（ESV）]等指标。组间比较采用方差分析和秩和检验，心功能的影响因素采用多元线性回归分析。 结果:共入组患者142例，其中男77例，女65例，年龄51（42，59）岁；失访3例。分为肥胖组（48例）、超重组（45例）及正常体重组（46例）。（1）三组间比较BMI、腰围、腰臀比、VAT质量、VAT面积、右室前壁、心尖脂肪差异有统计学意义（均 P<0.05）。（2）单因素分析显示，射血分数与BMI、腰围、VAT面积、右室前壁脂肪、心尖脂肪相关（β=-0.590~-0.023，均 P<0.05），EDV与VAT面积、心尖脂肪相关（β=0.196、4.357，均 P<0.05），ESV与VAT面积、心尖脂肪相关（β=0.113、2.675，均 P<0.05）。（3）多元线性回归显示，在模型Ⅰ（未调整）和模型Ⅱ（调整年龄和性别）中，心尖脂肪每增加1 mm，射血分数分别下降0.590%、0.645%，EDV依次增加4.357%、4.835%，ESV依次增加2.675%、2.931%。 结论:BMI、腰围、VAT质量、VAT面积、右室前壁脂肪、心尖脂肪是影响心功能的重要影响因素，尤以心尖脂肪对心功能的影响最大。

目的:调查某铀矿周围与对照区（察布查尔县和乌鲁木齐市）饮用水的放射性水平并进行健康风险评估。方法:完全随机采集2020—2022年某铀矿周围（51份）和对照区[（察布查尔县（73份）和乌鲁木齐市（52份）]的饮用水样品共176份，测定其总α和总β放射性水平及放射性核素 238U、 232Th、 226Ra、 40K的浓度。比较不同区域（铀矿周围、察布查尔县和乌鲁木齐市）、不同水源来源（地表水、地表水＋地下水、地下水）、距铀矿不同距离（5、10、15、20、25 km）的饮用水的总放射性水平。采用美国国家环境保护局（USEPA）、国际放射防护委员会和WHO推荐的方法分别估算饮用铀矿周围和对照区饮用水所致成年人的年有效剂量；采用USEPA提出的致癌风险因子评估居民终身致癌风险。对不同区域、不同水源、距铀矿不同距离的饮用水样品的总放射性水平进行单因素方差分析，若分析显示差异存在统计学意义时，进一步采用最小显著性差异法进行多重比较。 结果:不同区域饮用水样品中总α和总β放射性水平差异均有统计学意义[（0.13±0.04）Bq/L对（0.12±0.08）Bq/L对（0.08±0.03）Bq/L， F=9.854， P< 0.01]、[（0.17±0.06）Bq/L对（0.13±0.10）Bq/L对（0.10±0.03）Bq/L， F=10.522， P<0.01]。不同水源来源的饮用水样品中总α和总β放射性水平差异均无统计学意义（ F=2.849、1.352，均 P>0.05）。距铀矿不同距离的饮用水样品中总α和总β放射性水平差异均有统计学意义[（0.16± 0.02）Bq/L对（0.07±0.02）Bq/L对（0.11±0.02）Bq/L对（0.15±0.03）Bq/L对（0.14±0.03）Bq/L， F= 21.720， P<0.01]、[（0.24±0.04）Bq/L对（0.09±0.01）Bq/L对（0.19±0.01）Bq/L对（0.17±0.03）Bq/L对（0.13±0.04）Bq/L， F=46.364， P<0.01）。距铀矿5 km处的饮用水样品中总α和总β放射性水平均值均最高。176份饮用水样品的总α放射性水平均值为（0.11±0.06）Bq/L，0.5%的饮用水超过国家标准限值（0.5 Bq/L）。通过饮用水样品中总α放射性水平估算饮用该饮用水所致年有效剂量平均值为（0.040±0.003）mSv/年。通过饮用水样品中放射性核素 238U、 232Th、 226Ra、 40K浓度估算饮用该饮用水所致年有效剂量平均值分别为（0.030±0.004）、（0.018±0.003）、（0.084±0.04）、（0.005±0.003）mSv/年。饮用该饮用水所致居民终身致癌风险为1.75×10 -13~4.35×10 -11。 结论:某铀矿周围和对照区（察布查尔县和乌鲁木齐市）的饮用水放射性水平较低，饮用该区域饮用水所致年有效剂量和终身致癌风险较低，不会造成可观测到的不良健康效应，对周围居民所造成的健康风险较低。