目的： 探讨外源性硫化氢供体GYY4137对D半乳糖（DGal）诱导的衰老骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs）成骨能力的影响及潜在机制。方法： 应用不同浓度的DGal诱导BMSCs的衰老，CCK8法检测细胞活性及β半乳糖苷酶染色检测细胞衰老的程度，确定DGal的最佳诱导浓度；应用不同浓度GYY4137进行干预后，检测GYY4137对正常BMSCs及衰老BMSCs细胞活性的影响，确定GYY4137最佳干预浓度。实验设CON组、GYY4137组、DGal组、DGal+GYY4137组，作相应处理后对各组BMSCs进行β半乳糖苷酶染色，qRTPCR法检测衰老相关基因（P16、P21、P53）和衰老相关炎症因子（IL6、IL8、TNFα）的mRNA相对表达。对各组细胞进行成骨诱导分化，qRTPCR法检测成骨分化相关因子碱性磷酸酶（ALP）、骨钙素（OCN）、Runt相关转录因子2（RUNX2)、人骨形态发生蛋白2（BMP2）的mRNA表达，通过茜素红染色法检测钙结节形成数量，ALP染色法检测ALP的生成。通过转录组测序检测DGal组、DGal+GYY4137组两组细胞的差异表达基因，根据测序结果，qRTPCR对差异基因进行验证。应用Dickkopf相关蛋白1（dickkopf Wnt signaling pathway inhibitor 1，DKK1）抑制Wnt10b的表达后，检测各组衰老BMSCs的成骨分化能力。结果： DGal的最佳诱导浓度为30 g/L，GYY4137的最佳干预浓度为10 μmol/L，DGal组β半乳糖苷酶阳性细胞比例显著增加，衰老相关基因（P16、P21、P53）及衰老相关炎症因子（IL6、IL8、TNFα）mRNA的表达量显著升高（P＜005或P＜001），应用GYY4137干预后，以上细胞衰老相关指标显著降低（P＜005或P＜001）。在成骨分化过程中，DGal+GYY4137组ALP、OCN、RUNX2、BMP2的mRNA表达，钙结节阳性比例显著高于DGal组（P＜005或P＜001）；测序结果发现Wnt/βcatenin信号通路中的Wnt10b表达有显著差异，qRTPCR结果显示GYY4137可增加衰老细胞Wnt10b及其下游基因βcatenin的表达（P＜005或P＜001），与测序结果一致。应用100 μg/L DKK1抑制Wnt10b的表达后，GYY4137处理的衰老BMSCs成骨分化相关基因ALP、OCN、RUNX2、BMP2的mRNA表达明显降低（P＜005或P＜001）。结论： GYY4137可以缓解DGal诱导的BMSCs衰老进程，并且可能通过Wnt/βcantenin信号通路促进衰老BMSCs的成骨分化。

Objective:To explore the potential mechanism of Astragalus membranaceus and Atractylodes macrocephala in the treatment of osteoporotic fracture (OPF) based on network pharmacology.  Methods:  The active components and action targets of Astragalus membranaceus and Atractylodes macrocephala were screened by pharmacologic database and Analysis Platform (TCMSP) . GeneCards, OMIM and TTD databases related to disease targets were retrieved to screen the action targets of OPF, and then the intersection core targets of Astragalus membranaceus, Atractylodes macrocephala membranaceus and OPF were screened.  Cytoscape 3.7.1 software was used to construct a compoundtargetdisease network, and DAVID database was used to analyze the enrichment of GO function and KEGG pathway.  Results: Based on TCMSP database, 19 active components and 115 intersection targets of Astragalus membranaceus-Atractylodes for OPF were screened. A total of 568 items were obtained by GO functional enrichment analysis, including 433 biological process items, 43 cell composition items and 92 molecular function items.  KEGG pathway enrichment analysis revealed 111 items, including AGE-RAGE, IL-17, TNF, prostate cancer, Toll-like receptor, HIF-1, NF-κB signaling pathway, etc.  Conclusion: Astragalus membranaceus and Atelectylodes atelectylodes may regulate inflammatory response and internal balance of osteoclasts by regulating TNF, NF-κB and HIF-1 signaling pathways, and thus exert therapeutic actions on OPF.

Objective:To evaluate the clinical efficacy of the sequential infusion of bone cement when percutaneous kyphoplasty(PKP) was conducted for the treatment of osteoporotic vertebral compression fracture(OVCF) with posterior wall bone defects. Methods: Patients with posterior wall bone defects OVCF treated in our department from May 2016 to July 2020 were retrospectively analyzed. A total of 32 patients meeting the inclusion and exclusion criteria were treated by the sequential infusion of bone cement when PKP was applied. The visual analog score(VAS), Oswestry disability index(ODI), Cobb angle and height of injured vertebra were recorded and compared before surgery and 1 and 3 months after surgery. Postoperative bone cement leakage and complications were also recorded. Results:The postoperative VAS,ODI and Cobb angle of injured vertebra were significantly lower than those before surgery(P＜0.05).The postoperative height of the injured vertebra was significantly higher than that before surgery(P＜0.05). However, there was no significant differences between followup results at 1 and 3 months postoperatively(P＞0.05). There were 3 cases (9.38%) of bone cement leakage, without any cases of spinal canal leakage. No surgical complications such as nerve injury and pulmonary embolism were observed. Conclusion:Sequential infusion of bone cement during PKP is a safe and effective surgery for the OVCF with posterior wall bone defects, which could effectively reduce the bone cement leakage.

Objective： To study the effects of bone marrow mesenchymal stem cells （BMMSCs） infusion on plasma C3 and IgG concentrations， and CXCR4 expression of B cells and plasma cells in MRL/lpr lupus mice， and to explore the mechanism of BMMSCs in regulating disease activity of systemic lupus erythematosus. Methods： BMMSCs were isolated from C57BL/6 mice and cultured， and then injected into MRL/lpr lupus mice by tail transfusion. All mice were divided into C57BL/6 group （C57BL/6 mice were not injected ）， MRL/lpr group （MRL/lpr mice were not injected with BMMSCs）， BMMSCs infusion group （MRL/lpr mice were injected with BMMSCs at 12 weeks）， and PBS control group （MRL/ lpr mice were injected with PBS at 12 weeks）.At the end of the 20th week， peripheral blood， bone marrow and spleen of mice were taken for flow cytometry analysis to detect CXCR4 expression in B and plasma cells， and then the plasma of the 4 groups were taken for ELISA to detect IgG and C3 level. The kidney of mice was stained with hematoxylineosin. Results： ① Compared with the C57BL/6 group， the plasma C3 level were decreased and IgG level were significantly increased in the MRL/lpr group（all P<0.01）. Compared with C57BL/6 group， the expression of CXCR4 on B cell surface in MRL/lpr group was significantly decreased in peripheral blood， and was significantly increased in bone marrow and spleen（all P<0.01）. The proportion of CXCR4 on plasma cell surface in bone marrow（P<0.05） and spleen（P<0.01） of MRL/lpr group was significantly higher than that of C57BL/6 mice.② Compared with PBS or MRL/lpr group， the level of C3 in BMMSCs group was significantly increased， while the level of IgG was significantly decreased（all P<0.01）. Compared with the PBS group， the expression of CXCR4 on B cell surface of BMMSCs group was significantly increased in peripheral blood（P<0.01）， and significantly decreased in bone marrow（P<0.01） and spleen（P<0.05）. Compared with the MRL/lpr group， the expression of CXCR4 on B cell surface of bone marrow and spleen was significantly decreased （all P<0.01）.③ Compared with PBS group or MRL/lpr group， the expression of CXCR4 on plasma cell surface in bone marrow and spleen of BMMSCs group was significantly decreased（all P<0.05）.④ Renal HE staining suggested attenuation of renal inflammation in MRL/lpr mice after infusion of BMMSCs. Conclusion： BMMSCs infusion can regulate the expression of CXCR4 on the surface of B and plasma cells and improve the disease activity index of MRL/lpr mice.

Objective: To analyse the changes of selfreferential network （SRN）and cerebrospinal fluid pathological markers amyloid βprotein （Aβ）, total tau （ttau） and phosphorylated tau （p-tau） for patients with amnestic mild cognitive impairment（aMCI）. Methods: A total of 51 aMCI patients were enrolled in the study. According to the cognitive awareness index, they were divided into the aMCI group with impaired cognitive awareness（n=20）and the aMCI group（n=31）with cognitive awareness preserved. All patients underwent resting functional magnetic resonance （rsfMRI）, lumbar puncture, and cognitive assessment. Independent component analysis was used to extract SRN; two-sample t test was used to explore the functional connectivity of SRN and cerebrospinal fluid （Aβ, ttau and ptau）changes; Pearson correlation analysis was used to explore the relationship between cognitive awareness index and functional connectivity changes and the levels of cerebrospinal fluid Aβ, t-tau and p-tau. Results: The aMCI group with impaired cognitive awareness showed hyperconnectivity in the bilateral medial orbitofrontal cortex, left anterior cingulate gyrus and right rectus gyrus （AlphaSim correction, P<0.001, voxel number>17）. The difference in brain function connectivity between the two groups was negatively correlated with the cognitive awareness index（r=-0.457, P<0.000）. The level of Aβ in cerebrospinal fluid in the aMCI group with impaired cognitive awareness was significantly lower than that of the aMCI group with cognitive awareness preserved（P<0.01）, ptau and ttau in cerebrospinal fluid were significantly increased than those of the aMCI group with cognitive awareness preserved（P<0.001）. The cerebrospinal fluid Aβ level was positively correlated with cognitive awareness index（r=0.331, P=0.025）, and cerebrospinal fluid t-tau and p-tau levels were negatively correlated with cognitive consciousness index （r=-0.534,-0.508, both P<0.001）. Conclusion: SRN is the neuroimaging basis of cognitive awareness; aMCI patients with impaired cognitive awareness have impaired functional connectivity of SRN and more severe deposition of Alzheimer′s disease pathological markers.

目的:  探讨中性粒细胞明胶酶相关脂质运载蛋白（NGAL）、尾加压素Ⅱ和胱抑素C联合检测对早期糖尿病肾病（DKD）的诊断价值。方法: 选取我院住院T2DM患者60例，以24 h尿微量白蛋白（mAlb）含量分为单纯T2DM组（mAlb<30 mg/24 h）和早期DKD组(mAlb在30～300 mg/24 h）。选择同期健康体检者30例作为健康对照组。AU5800生化仪检测各组空腹血清NGAL、胱抑素C和尿肌酐，免疫比浊法检测血清尾加压素Ⅱ和尿mAlb。结果：  与健康对照组相比较，单纯T2DM组估算肾小球滤过率（eGFR）、尿白蛋白与肌酐比值（UACR）、NGAL、尾加压素Ⅱ和胱抑素C差异无统计意义；早期DKD组除eGFR显著降低外，UACR、NGAL、尾加压素Ⅱ和胱抑素C均显著高于单纯T2DM组和健康对照组（P均<0.05）。早期DKD组中，NGAL、尾加压素Ⅱ和胱抑素C含量均与UACR呈正相关（r值分别为0.651、0.682、0.723， P均<0.05）。血清NGAL、尾加压素Ⅱ和胱抑素C联合检测DKD的ROC曲线下面积、敏感性和特异性分别为0.937、92.40%和86.30%，显著高于单项检测的诊断价值（P<0.05）。结论： 在DKD早期诊断中，血清NGAL、尾加压素Ⅱ和胱抑素C联合检测优于单项检测。