Matrix gamma-carboxyglutamic acid protein is a key regulator of PTH-mediated inhibition of mineralization in MC3T3-E1 osteoblast-like cells

As part of its overall function as a major regulator of calcium homeostasis, PTH stimulates bone resorption and inhibits osteoblast-mediated biomineralization. To determine the basis for the inhibitory actions of this hormone, we compared the time course of PTH-dependent inhibition of mineralization in MC3T3-E1 osteoblast-like cells with changes in mRNA levels for several extracellular matrix proteins previously associated either with induction or inhibition of mineralization. Mineralizing activity was rapidly lost in PTH-treated cells ( approximately 30% inhibition after 3 h, 50% inhibition at 6 h). Of the proteins examined, changes in matrix gamma-carboxyglutamic acid protein were best correlated with PTH-dependent inhibition of mineralization. Matrix gamma-carboxyglutamic acid protein mRNA was rapidly induced 3 h after PTH treatment, with a 6- to 8-fold induction seen after 6 h. Local in vivo injection of PTH over the calvaria of mice also induced a 2-fold increase in matrix gamma-carboxyglutamic acid protein mRNA. Warfarin, an inhibitor of matrix gamma-carboxyglutamic acid protein gamma-carboxylation, reversed the effects of PTH on mineralization in MC3T3-E1 cells, whereas vitamin K enhanced PTH activity, as would be expected if a gamma-carboxyglutamic acid-containing protein were required for PTH activity. Levels of the other mRNAs examined were not well correlated with the observed changes in mineralization. Osteopontin, an in vitro inhibitor of mineralization, was induced approximately 4-fold 12 h after PTH addition. Bone sialoprotein mRNA, which encodes an extracellular matrix component most frequently associated with mineral induction, was inhibited by 50% after 12 h of PTH treatment. Osteocalcin mRNA, encoding the other known gamma-carboxyglutamic acid protein in bone, was also inhibited by PTH, but, again, with a significantly slower time course than was seen for mineral inhibition. Taken together, these results show that the rapid inhibition of osteoblast mineralization induced by in vitro PTH treatment is at least in part explained by induction of matrix gamma-carboxyglutamic acid protein.     

Postnatal maturational changes in rat pelvic autonomic ganglion cells: a mixture of steroid-dependent and -independent effects

Androgens have potent effects on the maturation and maintenance of a number of neural pathways involved in reproductive behaviors in males. Most studies in this area have focused on central pathways, but androgen receptors are expressed by many peripheral neurons innervating reproductive organs, and previous studies have demonstrated structural and chemical changes in these neurons at puberty and after castration. We have performed the first electrophysiological comparison of pelvic autonomic ganglion neurons in male rats before and after puberty and following pre- or postpubertal castration. Studies were performed in vitro on intact ganglia with hypogastric and pelvic nerves attached to allow synaptic activation of sympathetic or parasympathetic neurons, respectively. Pelvic ganglion neurons underwent many changes in their passive and active membrane properties over the pubertal period, and some of these changes were dependent on exposure to circulating androgens. The most pronounced steroid-dependent effects were on membrane capacitance (soma size) in sympathetic neurons and duration of the action potential afterhyperpolarization in tonic neurons. Our study also showed that rat pelvic ganglion cells and their synaptic inputs were more diverse than previously reported. In conclusion, this study demonstrated that rat pelvic ganglion neurons undergo considerable postnatal changes in their electrophysiological properties. The steroid dependence of some of these changes indicates that circulating androgens may influence reproductive behaviors at many locations within the nervous system not just in the brain and spinal cord.     

Ethanol-wet bonding may improve root dentine bonding performance of hydrophobic adhesive

Objectives

The current study aimed to assess ethanol-wet dentine surfaces by atomic force microscopy (AFM), and to evaluate the efficacy of ethanol-wet bonding on root dentine by determining the shear bond strength (SBS) and interfacial nanoleakage expression.

Methods

Flat dentine slices from human premolar roots were randomly grouped into five. All specimens were acid-etched, rinsed, and left moist. They were then treated with 100% ethanol for 0 s (control group), 20 s (Group 1), 60 s (Group 2), three 60 s periods (Group 3), or stepwise ethanol application (Group 4). After treatment, each group was bonded either with Adper™ Scotchbond™ Multi-Purpose (Scotchbond) or experimental hydrophobic adhesive. Nano-scale adhesion forces (Fad) were probed by AFM and analysed using one-way ANOVA. The SBS results were analysed using two-way ANOVA. Tukey's test was employed for multiple comparisons.

Results

Ethanol-wet protocols significantly decreased the value of Fad (p < 0.001). When bonded with Scotchbond, ethanol treatment did not affect the bond strength (p > 0.05), but decreased the interfacial nanoleakage. The SBS values of the groups bonded with hydrophobic adhesive varied with different ethanol-wet protocols (p < 0.05). Decreased nanoleakage was manifested in all experimental groups, except Group 1. Compared with the classical water-wet bonding with Scotchbond in the control group, Group 4 bonded with hydrophobic adhesive exhibited a significantly higher bond strength (p < 0.05).

Conclusions

Ethanol-wet bonding using a stepwise ethanol application protocol may have potential benefits in the root dentine bonding of hydrophobic adhesive.     

Biological Changes on the Compressing Side of Orthodontic Teeth Stimulating by Soft laser

目的：研究在弱激光作用下接受正畸力作用的牙齿的压力侧出现的生物学变化，为临床应用弱激光加速牙齿移动提供理论依据。方法：实验动物分2组，每组20只。A组动物接受正畸力，B组动物除接受正畸力之外，还接受弱激光照射。采用免疫组化检测层连蛋白的组间表达变化，采用原位杂交检测RANKL(receptor activator of NF-kB ligand)mRNA的组间表达变化。结果：免疫组化结果表明，新生血管活跃的高峰期出现在接受正畸力的第7d。与A组相比，接受弱激光照射的B组呈现层连蛋白的高表达。同样，与A组相比，原位杂交检测发现在接受弱激光照射的B组中，正畸牙压力侧呈现RANKL mRNA的高表达。结论：弱激光照射后能够促进正畸牙压力侧血管新生(呈现层连蛋白高表达)，从而促进破骨细胞的分化激活。此外弱激光还能够促进破骨细胞活化因子RANKL的表达，增强正畸牙压力侧的破骨细胞的活性，加快骨改建。     

膝关节置换后注射含类固醇鸡尾酒治疗安全性的荟萃分析

背景:类固醇具有强大的抗炎、止吐及镇痛作用而被广泛用于围术期镇痛,有研究表明膝关节置换后膝关节周围注射含类固醇的鸡尾酒镇痛疗法可缓解术后疼痛、改善膝关节活动度及减少相关并发症,但也有研究认为类固醇可增加术后感染、髌腱断裂等风险。因此膝关节周围注射含类固醇鸡尾酒疗法在膝关节置换后镇痛中的安全性和疗效尚存在争议。目的:通过荟萃分析系统评价膝关节置换后运用含类固醇鸡尾酒疗法镇痛的疗效和安全性。方法:检索PubMed/Medline、Cochrane Central Register of Controlled Trials、EMBASE数据库2019年4月前的文献,收集所有膝关节置换后局部运用类固醇镇痛的随机对照试验,筛选出符合条件的文献。由2名研究者分别单独使用Cochrane 5.0风险偏倚评估工具对文献方法学偏倚进行评价,提取文献数据并用Review Manager 5.2软件进行荟萃分析。结果与结论:①共纳入10篇随机对照试验,820例患者;②Meta分析显示:类固醇组术后第1天的目测类比评分低于对照组[MD=-1.52,95%CI(-2.94,-0.10),P=0.04],术后第1,2,3,4,5天的关节活动度高于对照组[MD=11.57,95%CI(9.85,13.30),P<0.000 01;MD=9.03,95%CI(6.67,11.38),P<0.000 01:MD=5.73,95%CI(0.85,10.60),P=0.02:MD=5.53,95%CI(0.68.10.38),P=0.03;MD=5.90.95%CI(0.87.10.93),P=0.02],吗啡使用量少于对照组[MD=-7.94,95%CI(-14.35,-1.53),P=0.02],住院时间短于对照组[MD=-0.98,95%CI(-1.25,-0.71),P<0.000 01],直腿抬高所需时间短于对照组[MD=0.65,95%CI(-0.86,0.44),P<0.000 01],术后C-反应蛋白水平低于对照组[WMD=-4.82,95%CI(7.41,2.23),P=0.000 3];两组膝关节KSS评分、并发症发生率比较差异无显著意义;③结果表明,膝关节置换后关节周围运用含类固醇的鸡尾酒疗法安全有效。     

内镜下分段切除儿童巨大肠息肉的临床疗效探讨

目的 该文旨在探讨内镜下分段切除儿童巨大肠息肉的临床疗效及安全性.方法 回顾性分析该院消化内科和普外科收治的66例巨大肠息肉患儿,43例采取内镜下分段切除治疗,23例行外科手术.观察两组患儿禁食时间、住院时间及不良反应发生情况.结果 两组患儿均成功进行息肉切除术,内镜治疗组在禁食时间、住院时间及住院费用上均优于外科手术...     

FGF and ERK signaling coordinately regulate mineralization-related genes and play essential roles in osteocyte differentiation

To examine the roles of FGF and ERK MAPK signaling in osteocyte differentiation and function, we performed microarray analyses using the osteocyte cell line MLO-Y4. This experiment identified a number of mineralization-related genes that were regulated by FGF2 in an ERK MAPK-dependent manner. Real-time PCR analysis indicated that FGF2 upregulates Ank, Enpp1, Mgp, Slc20a1, and Dmp1 in MLO-Y4 cells. Consistent with this observation, the selective FGF receptor inhibitor PD173074 decreased Ank, Enpp1, Slc20a1, and Dmp1 mRNA expression in mouse calvaria in organ culture. Since Dmp1 plays a central role in osteocyte differentiation and mineral homeostasis, we further analyzed FGF regulation of Dmp1. Similar to FGF2, FGF23 upregulated Dmp1 expression in MLO-Y4 cells in the presence of Klotho. Furthermore, increased extracellular phosphate levels partially inhibited FGF2-induced upregulation of Dmp1 mRNA expression, suggesting a coordinated regulation of Dmp1 expression by FGF signaling and extracellular phosphate. In MLO-Y4 osteocytes and in MC3T3E1 and primary calvaria osteoblasts, U0126 strongly inhibited both basal expression of Dmp1 mRNA and FGF2-induced upregulation. Consistent with the in vitro observations, real-time PCR and immunohistochemical analysis showed a strong decrease in Dmp1 expression in the skeletal elements of ERK1 ^−/− ; ERK2 ^flox/flox ; Prx1-Cre mice. Furthermore, scanning electron microscopic analysis revealed that no osteocytes with characteristic dendritic processes develop in the limbs of ERK1 ^−/− ; ERK2 ^flox/flox ; Prx1-Cre mice. Collectively, our observations indicate that FGF signaling coordinately regulates mineralization-related genes in the osteoblast lineage and that ERK signaling is essential for Dmp1 expression and osteocyte differentiation.     

Synthesis and preliminary evaluation in vitro of novel naproxen-dendritic peptide conjugates

A series of bone-targeting prodrugs, dendritic L-Asp and L-Glu peptides Naproxen conjugates have been synthesized in a convergent approach and well characterized by NMR and MS techniques. Their solubility in water and hydroxyapatite affinity were evaluated in in vitro conditions. All the prodrugs were water soluble and exhibited high affinity to HAP. Compound NAP-G₂-Asp was found more potent in HAP binding. The efficient release of the active drug moiety (naproxen) occurred by the cleavage of an amide bond in physiological conditions. These results indicated that the dendritic peptides might become a delivery system for bone tissues and provided an effective entry to the development of new bone-targeting molecules.     

巴马神酒调节免疫及抗脂质过氧化药理作用研究

摘 要 目的： 观察巴马神酒对小鼠免疫功能、肝脏脂质过氧化损伤及游泳耐力的药理作用。方法: 采用碳粒廓清实验法及环磷酰胺致小鼠免疫低下模型评价巴马神酒对免疫功能的作用;建立四氯化碳致小鼠急性肝损伤模型,以血清中丙氨酸氨基转移酶（ALT）、天门冬氨酸转移酶（AST）、超氧化物歧化酶（SOD）以及丙二醛（MDA）的水平,评价巴马神酒抗肝脏脂质过氧化损伤作用;以小鼠负重游泳的持续时间,评价巴马神酒提高游泳耐力的功能。结果: 巴马神酒能提高小鼠吞噬指数K及吞噬系数α（与空白组比较,P<0.01或P<0.05）;对环磷酰胺诱发的胸腺、脾脏脏器指数的下降有一定的提高（与模型组比较,P<0.05）;能够降低四氯化碳致小鼠急性肝损伤血清中AST、ALT含量,降低血清MDA的含量,增强SOD的活性（与模型组比较,P<0.01或P<0.05）;可以延长小鼠负重游泳时间（与空白组比较,P<0.01或P<0.05）。结论: 巴马神酒具有提升小鼠巨噬细胞吞噬能力、保护免疫低下小鼠的胸腺和脾脏指数的作用,并具有抗肝脏脂质过氧化损伤和提高游泳耐力的作用。     

Clinicopathologic features and clinical outcomes of intravenous leiomyomatosis of the uterus: A case series

Rationale:Intravenous leiomyomatosis (IVL) is a rare and special type of smooth muscle tumor originating in the uterus. It is classified as a benign disease according to its histological features but shows the behavioral characteristics of a malignant tumor. It is easily misdiagnosed and recurrent. The purpose of this study was to retrospectively analyze clinicopathological data of 25 cases of IVL in order to enhance clinicians’ understanding of this rare disease.Patient concerns:We screened and identified 25 cases of IVL at our hospital from October 2013 to January 2020. Five patients had tumors.Diagnoses:The diagnosis in each case was pathologically confirmed after surgical treatment.Interventions:All patients were managed surgically. Although the surgical procedures were different, the surgical approach was geared towards achieving complete excision. Three patients received hormonal therapy with gonadotropinreleasing hormone agonists after surgery.Outcomes:We retrospectively reviewed all medical records and analyzed the clinicopathologic features and clinical outcomes of this disease as well as the correlations between the clinical features and risk of recurrence. Neither the symptoms nor the preoperative imaging results were suggestive of IVL in any of the cases. Except for two patients who were lost to follow-up, twenty-three patients who were followed up are still alive. Three patients experienced a recurrence.Lessons:The clinical manifestations and ultrasound images of IVL in the early stages are not typical; thus, IVL is easily misdiagnosed as uterine leiomyoma. Radiologists, pathologists, and surgeons should have a thorough understanding of IVL and a high index of vigilance for IVL in clinical practice. Surgery should always be aimed at achieving complete tumor excision. Patients with large lesions (≥7 cm) and lesions extending to the broad ligament may have an increased risk of recurrence. Early detection, diagnosis, and treatment are very important; once the diagnosis is confirmed, regular follow-ups are crucial.