Mineralized nodule formation by cultures of human dental pulp-derived fibroblasts

Pulp fibroblasts were isolated from human deciduous and supernumerary teeth and cultured in vitro. With continued culture in normal tissue-culture medium, six pulp fibroblast strains formed cell nodules after 10–15 days. By electron microscopy the nodules had matrix vesicles, and needle-shaped crystals associated with a dense network of collagen fibrils. The crystalline material exhibited a pattern consistent with hydroxyapatite when nodules were examined by X-ray diffractometry. Furthermore, the cells showed high levels of alkaline phosphatase activity, which could be increased more than seven-fold by the addition of 1,25(OH)₂D₃ (5 × 10⁻⁹−5 × 10⁻⁸M). In addition to the production of type I collagen, these cells also synthesized fibronectin and osteonectin. The formation of mineralized tissue nodules by pulp cells in vitro provides a useful system for study of the pathological calcification of pulp tissues.     

In situ localization of tenascin mRNA in developing mouse teeth

Tenascin is a large extracellular-matrix glycoprotein found in developing connective tissue. A cDNA probe to mouse tenascin, mTN2, was used to determine the cellular origins of this molecule in the murine tooth germ by in situ hybridization. At embryonic day 19, a hybridization signal significantly greater than background was detected with mTN2 in the subodontoblastic layer of the dental mesenchyme and in the inner enamel epithelium of the enamel organ. At postnatal day 1, a signal was detected over pre-odontoblasts and the strata intermedium and externum. No tenascin mRNA was detected in odontoblasts or the stellate reticulum at either age, and hybridization in ameloblasts was not significantly greater than background at postnatal day 1. Thus, much of the tenascin found throughout developing teeth appears to be synthesized by pre-odontoblasts and the inner enamel epithelium, the two populations of cells destined to generate mineralized matrix.     

Immunohistochemical localization of very late activation integrins in healthy and diseased human gingiva

The β1-integrins (VLA family) are cellular adhesion molecules (CAM) that play a major role in cell-cell and cell-matrix interactions. The expression pattern of CAM was studied in 5 clinically normal volunteers with healthy gingiva and in 18 patients with clinically different stages of periodontitis. In healthy human gingiva α2. α3 and α6 integrin chains were found in a characteristic distribution, showing a broad continuous expression on the junctional and sulcular epithelium sites. The expression of these integrins was demonstrated primarily on the basal cell layers and in some cells of the stratum spinosum. Inflammatory stages of periodontitis revealed further upregulation of α2, α3 and α6 integrins into the junctional and sulcular epithelial cells, which correlated with the stage of the periodontitis and the extent of the cellular infiltration. α4 and α6 were found to be the predominant β1 integrin chains on inflammatory cells. The amount of δ4 and ş6 positive infiltrative cells increased with the number of inflammatory cells. VCAM-1. the corresponding cell-cell ligand of VLA-4 (α4) was present on the majority of subepithelial vessels in all stages of gingivitis and periodontitis. The α5 subunit was expressed on both endothelium and gingival connective tissue cells. Samples from advanced periodontitis cases showed a higher number of a5 positive mononuclear cells. In comparison to normal epidermis, a human gingival epithelial cells express higher levels of integrins. This expression is further upregulated in advanced stages of periodontitis, indicating changes of the β1 integrin organization.     

Involvement of Nrf2, JAK and COX pathways in acetaminophen-induced nephropathy: Role of some antioxidants

ObjectivesAcetaminophen (APAP)-induced nephrotoxicity is detrimental consequence for which there has not been a standardized therapeutic regimen. Although, N-acetylcysteine (NAC) is a well-known antidote used in APAP-induced hepatotoxicity, its benefit in nephrotoxicity caused by APAP is almost lacking. This study aimed to compare the possible protective effect of thymoquinone (TQ), curcumin (CR), and α-lipoic acid (α-LA), either in solo or in combination regimens with that of NAC against APAP-induced renal injury.Design and methodRats were divided into nine groups; control group, APAP intoxicated group (1000 mg/kg; orally), and the remaining seven groups received, in addition to APAP, oral doses of NAC, TQ, CR, α-LA, CR plus TQ, TQ plus α-LA, or CR plus α-LA. The first dose of the aforementioned antioxidants was given 24 h before APAP, and then the second dose was given 2 h after APAP, whereas the last dose was given 10 h after administration of APAP.ResultsTreatment with APAP elevated kidney markers like serum uric acid, urea, and creatinine. In addition, it increased the serum level of tumor necrosis factor alpha (TNF-α), interleukin-1beta (IL-1β) and thiobarbituric acid reactive species (TBARS). Also, the protein expression of renal janus kinase (JAK) and cyclooxygenase (COX)-2 were all upregulated by APAP. In contrast, the expression of Nrf2 and the renal levels of superoxide dismutase and glutathione were downregulated. Treatment with the indicated natural antioxidants resulted in amelioration of the aberrated parameters through exhibiting anti-inflammatory, antioxidant and free radical-scavenging effects with a variable degree.ConclusionThe combined administration of CR and TQ exerted the most potent protection against APAP-induced nephrotoxicity through its anti-inflammatory and free radical-scavenging effects (antioxidant) which were comparable to that of NAC-treatment.     

液体负平衡应用于感染性休克患者对APACHEII评分的影响

摘要:目的 探讨液体负平衡对感染性休克患者急性生理学与慢性健康状态（APACHEII）评分的影响分析。方法 选取2018年1月～2020年1月本院收治的92例感染性休克患者,按不同液体管理模式分为常规组（采用液体平衡）和研究组（采用液体负平衡管理模式）各46例。对比分析2组患者心肺功能及APACHEII评分以及不良反应。结果 管理前2组患者全心舒张末期容量指数（GED-CVI）和血管外肺水指数（EVLWI）比较无统计学差异（P>0.05）,管理后2组GED-CVI、EVLWI水平均增加（P＜0.05）,研究组上述水平高于常规组（P＜0.05）;研究组和常规组均完成了管理计划,2组管理前APACHEII评分比较,差异均无统计学意义（P＞0.05）,管理后均低于管理前（P＜0.05）,管理后研究组低于常规组（P＜0.05）;2组患者均未见明显不良反应发生。结论 液体负平衡应用于感染性休克患者,可改善心肺功能指标,降低患者APACHEII评分,可作为临床管理及防控的参考指标。     

姜黄素调控miR-199a-3p基因抑制前列腺癌细胞增殖、迁移和侵袭的研究

目的探讨姜黄素调控miR-199a-3p的基因表达对前列腺癌C4-2细胞增殖、迁移和侵袭的影响。方法将miR-199a-3p抑制物及阴性对照转染至C4-2细胞中,并分别标记为anti-miR-199a-3p组和anti-miR-con组;将仅加入脂质体的C4-2细胞标记为对照组。运用MTT法检测通过姜黄素(0、20、40、80μmol/L)处理的C4-2细胞的增殖情况。40μmol/L姜黄素处理C4-2细胞后,Transwell法检测细胞的迁移和侵袭能力;qRT-PCR检测细胞的miR-199a-3p表达量。将inhibitor NC、miR-199a-3p inhibitor转染至C4-2细胞中,再用40μmol/L姜黄素处理48 h,采用MTT法、Transwell法检测各组细胞的增殖、迁移和侵袭情况;Western blot检测各组C4-2细胞中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、β-catenin、Cyclin D1和c-Myc的蛋白表达量。结果与对照组比较,姜黄素能明显抑制C4-2细胞的增殖、迁移和侵袭(P<0.05)。姜黄素(40μmol/L)处理后,C4-2细胞中miR-199a-3p的表达量显著增加(P<0.05)。抑制miR-199a-3p的表达,可逆转姜黄素对C4-2细胞增殖、迁移和侵袭的抑制作用(P<0.05)。使用姜黄素处理miR-199a-3p低表达的C4-2细胞后,与anti-miR-con组相比,anti-miR-199a-3p组C4-2细胞的MMP-2、MMP-9的表达量显著增加(P<0.05),β-catenin、Cyclin D1和c-Myc蛋白表达量均显著增加(P<0.05)。结论姜黄素可上调miR-199a-3p的表达,从而抑制前列腺癌C4-2细胞的增殖、迁移和侵袭。     

通督调神针刺法结合认知疗法干预卒中后抑郁患者临床研究

摘要：目的 观察通督调神针刺法结合认知疗法干预卒中后抑郁的临床效应,并探讨其疗效可能的作用机制。方法 将60例卒中后抑郁患者随机分为观察组和西药组,每组30例,两组在予以脑血管疾病基础治疗的同时,采用不同方法干预,观察组采用通督调神针刺法结合认知疗法治疗,西药组予以盐酸帕罗西汀片口服,疗程均为4周,治疗前后进行汉密尔顿抑郁量表(HAMD)、日常生活能力评定（Barthel指数）及副反应量表( TESS) 评估,观察两组临床疗效,测定患者血清 5-HT水平。结果 同组治疗后较治疗前比较,HAMD评分均显著降低(P<0.05),Barthel 指数均提高（P＜0.05）,组间比较,观察组优于西药组,差异具有统计学意义(P<0.05),治疗后两组血清 5-HT水平均较治疗前升高 (P<0.05) ,两组5-HT升高幅度比较无统计学意义(P>0.05),观察组总有效率为93. 3%,西药组为86.7%,两组比较差异具有统计学意义(P<0.05),观察组不良反应发生率较西药组发生率低,差异有统计学意义(P<0.05)。结论 通督调神针刺法结合认知疗法法干预卒中后抑郁,可以显著改善 PSD 患者的抑郁状态,其疗效机制可能与血清 5-HT水平上调有关,通督调神针刺法结合认知疗法是治疗卒中后抑郁行之有效、副反应低、操作重复性好的临床治疗手段。     

热毒宁注射液金银花提取过程轨迹研究

摘 要：目的：建立热毒宁注射液金银花提取过程轨迹,实现对提取过程的全程在线监控,为中药生产批次间质量控制提供参考。方法：收集多批次金银花提取过程的近红外光谱（NIRs）数据,并使用多元统计过程控制技术（MSPC）建立提取过程轨迹,实现对提取过程的全程监控。结果：建立的热毒宁注射液金银花提取过程的主成分得分,Hotelling T2和DModX轨迹图能够检测到过程的异常情况。结论：基于近红外光谱数据和MSPC技术建立的过程轨迹图可以反应中药生产的过程变化,可以用于对中药生产的全程监控。     

中医药快速抗抑郁研究进展

摘要：目的 观察通督调神针刺法结合认知疗法干预卒中后抑郁的临床效应,并探讨其疗效可能的作用机制。方法 将60例卒中后抑郁患者随机分为观察组和西药组,每组30例,两组在予以脑血管疾病基础治疗的同时,采用不同方法干预,观察组采用通督调神针刺法结合认知疗法治疗,西药组予以盐酸帕罗西汀片口服,疗程均为4周,治疗前后进行汉密尔顿抑郁量表(HAMD)、日常生活能力评定（Barthel指数）及副反应量表( TESS) 评估,观察两组临床疗效,测定患者血清 5-HT水平。结果 同组治疗后较治疗前比较,HAMD评分均显著降低(P<0.05),Barthel 指数均提高（P＜0.05）,组间比较,观察组优于西药组,差异具有统计学意义(P<0.05),治疗后两组血清 5-HT水平均较治疗前升高 (P<0.05) ,两组5-HT升高幅度比较无统计学意义(P>0.05),观察组总有效率为93. 3%,西药组为86.7%,两组比较差异具有统计学意义(P<0.05),观察组不良反应发生率较西药组发生率低,差异有统计学意义(P<0.05)。结论 通督调神针刺法结合认知疗法法干预卒中后抑郁,可以显著改善 PSD 患者的抑郁状态,其疗效机制可能与血清 5-HT水平上调有关,通督调神针刺法结合认知疗法是治疗卒中后抑郁行之有效、副反应低、操作重复性好的临床治疗手段。     

基于抑郁模型大鼠miR-665表达及靶基因功能分析探讨开心解郁方抗抑郁疗效机制

摘 要：目的：检测中药开心解郁方干预抑郁症模型大鼠过程中海马miR-665表达变化并对miR-665的靶基因进行生物信息分析,探讨miR-665在抑郁症的发病及开心解郁方抗抑郁机制中的作用。方法：建立慢性应激抑郁症大鼠模型并用开心解郁方干预42天,取海马组织提取总RNA,采用RT-qPCR检测各组大鼠海马miR-665相对表达量,采用TargetScan和microRNAorg数据库对miR-665进行靶基因预测,采用DAVID数据库对预测得到的靶基因进行GO功能富集分类及KEGG信号通路分析。结果：与正常组比较,模型组大鼠海马miR-665表达水平显著增高,差异具有统计学意义（P<0.01）;与模型组比较,中药组及西药组大鼠海马miR-665表达水平显著降低,差异具有统计学意义（P<0.05）。miR-665靶基因的生物学功能主要富集于有机物质反应等;信号通路主要富集于N-糖链的合成等。结论：miR-665可能参与了抑郁症病理机制过程调控,通过改善其表达异常可能为开心解郁方的抗抑郁机制之一,通过对其靶基因的功能预测分析对于后期继续研究开心解郁方干预miR-665的具体作用机制提供了方向和理论基础。