Fetal male lineage determination by analysis of Y-chromosome STR haplotype in maternal plasma

The aim of this study is to determine the fetus Y-STR haplotype in maternal plasma during pregnancy and estimate, non-invasively, if the alleged father and fetus belong to the same male lineage. The study enrolled couples with singleton pregnancies and known paternity. All participants signed informed consent and the local ethics committee approved the study. Peripheral blood was collected in EDTA tubes (mother) and in FTA paper (father). Maternal plasma DNA was extracted by using NucliSens EasyMAG. Fetal gender was determined by qPCR targeting DYS-14 in maternal plasma and it was also confirmed after the delivery. From all included volunteers, the first consecutive 20 mothers bearing male fetuses and 10 mothers bearing female fetuses were selected for the Y-STR analysis. The median gestational age was 12 weeks (range 12–36). All DNA samples were subjected to PCR amplification by PowerPlex Y23, ampFLSTR Yfiler, and two in-house multiplexes, which together accounts for 27 different Y-STR. The PCR products were detected with 3500 Genetic Analyzer and they were analyzed using GeneMapper-IDX. Fetuses’ haplotypes (Yfiler format) were compared to other 5328 Brazilian haplotypes available on Y-chromosome haplotypes reference database (YHRD). As a result, between 22 and 27 loci were successfully amplified from maternal plasma in all 20 cases of male fetuses. None of the women bearing female fetuses had a falsely amplified Y-STR haplotype. The haplotype detected in maternal plasma completely matched the alleged father haplotype in 16 out of the 20 cases. Four cases showed single mismatches and they did not configure exclusions; 1 case showed a mutation in the DYS 458 locus due to the loss of one repeat unit and 3 cases showed one DYS 385I/II locus dropout. All mismatches were confirmed after the delivery. Seventeen fetuses’ haplotypes were not found in YHRD and one of them had a mutation, which corresponded to the paternity probability of 99.9812% and 95.7028%, respectively. Three fetuses’ haplotypes occurred twice in YHRD, which corresponded to paternity probability of 99.9437%. In conclusion, high discriminatory fetal Y-STR haplotype could be determined from maternal plasma during pregnancy starting at 12 weeks of gestation. All male fetuses could be attributed to the alleged father male lineage early in pregnancy. The high probability of paternity associated with each case suggests that the relationship is not random and this strategy can be use as an alternative for male fetal kinship analysis.     

上颌唇侧倒置埋伏中切牙患者正畸矫治后埋伏牙发生及牙根吸收的影响因素分析

评估上颌唇侧倒置埋伏中切牙患者正畸矫治后埋伏牙发生及牙根吸收的影响因素。方法收集2019年3月-2021年3月于溧阳市人民医院就诊的1021例上颌唇侧倒置埋伏中切牙患者临床资料进行回顾性调查,查阅锥形束CT及X线图像。其中113例发现埋伏牙,按照是否出现牙根吸收分为牙根吸收组（51例）和牙根未吸收组（62例）。调取资料,包括性别、初诊年龄、平均体质量指数、埋伏牙位置、近远中分区、是否拔牙、正畸治疗时间、埋伏牙发生个数等。统计上述指标情况,分析正畸矫治后埋伏牙牙根吸收的单因素和多因素。结果 1021例患者中埋伏牙113例（11.07%）;牙根吸收组平均年龄、2~4区占比均低于牙根未吸收组（P ＜0.05）;牙根吸收组拔牙率、正畸治疗时间＞2年占比均高于牙根未吸收组（P ＜0.05）;两组性别、平均体质量指数、埋伏牙位置、病变位置、埋伏牙发生个数比较,差异无统计学意义（P ＞0.05）;Logistic结果显示,高龄、近远中分区（0~1区）、拔牙、正畸治疗时间＞2年均为正畸矫治后埋伏牙牙根吸收的高危因素（P ＜0.05）。结论 上颌唇侧倒置埋伏中切牙正畸矫治后埋伏牙牙根吸收与年龄、近远中分区、拔牙、正畸治疗时间有关,对于高危因素患者,临床应给予重视,采用早期干预措施,减少损伤。     

Increased Bone Resorption Is Associated With Increased Risk of Cardiovascular Events in Men: The MINOS Study

Better assessment of the association between cardiovascular disease and osteoporosis in older men may help identify shared etiologies for bone and heart health in this population. We assessed the association of BMD and bone turnover markers (BTMs) with risk of cardiovascular events (myocardial infarction or stroke) in 744 men ≥50 yr of age. During the 7.5‐yr prospective follow‐up, 43 strokes and 40 myocardial infarctions occurred in 79 men. After adjustment for confounders (age, weight, height, smoking, education, physical activity, self‐reported history of diabetes, hypertension, and prevalent ischemic heart disease), men in the lowest quartile of BMD at the spine, whole body, and forearm had a 2‐fold increased risk of cardiovascular events. Men in the highest quartile of bone resorption markers (deoxypyridinoline [DPD], C‐telopeptide of type I collagen) had a 2‐fold increased risk of cardiovascular events (e.g., multivariable‐adjusted hazard ratio [including additional adjustment for BMD] was 2.11 [95% CI: 1.26–3.56], for the highest quartile of free DPD relative to the lowest three quartiles). The results were similar for men without prevalent ischemic heart disease and for myocardial infarction and stroke analyzed separately. Our data suggest that men with low BMD or high bone resorption may be at increased risk of myocardial infarction and stroke in addition to fracture. Thus, men with osteoporosis may benefit from screening for cardiovascular disease. Further study to elucidate the biological mechanism shared by bone and vascular disease may help efforts to identify men at risk or develop treatment.     

Docetaxel inhibits bone resorption through suppression of osteoclast formation and function in different manners

Osteoclasts are formed from the monocyte-macrophage lineage in response to receptor activator of nuclear factor κB ligand (RANKL) expressed by osteoblasts. Bone is the most common site of breast cancer metastasis, and osteoclasts play roles in the metastasis. The taxane-derived compounds paclitaxel and docetaxel are used for the treatment of malignant diseases, including breast cancer. Here we explored the effects of docetaxel on osteoclastic bone resorption in mouse culture systems. Osteoclasts were formed within 6 days in cocultures of osteoblasts and bone marrow cells treated with 1,25-dihydroxyvitamin D₃ plus prostaglandin E₂. Docetaxel at 10⁻⁸ M inhibited osteoclast formation in the coculture when added for the entire culture period or for the first 3 days. Docetaxel, even at 10⁻⁶ M added for the final 3 days, failed to inhibit osteoclast formation. Osteoprotegerin, a decoy receptor of RANKL, completely inhibited osteoclast formation when added for the final 3 days. Docetaxel at 10⁻⁸ M inhibited the proliferation of osteoblasts and bone marrow cells. RANKL mRNA expression induced by 1,25-dihydroxyvitamin D₃ plus prostaglandin E₂ in osteoblasts was not affected by docetaxel even at 10⁻⁶ M. Docetaxel at 10⁻⁶ M, but not at 10⁻⁸ M, inhibited pit-forming activity of osteoclasts cultured on dentine. Actin ring formation and l-glutamate secretion by osteoclasts were also inhibited by docetaxel at 10⁻⁶ M. Thus, docetaxel inhibits bone resorption in two different manners: inhibition of osteoclast formation at 10⁻⁸ M and of osteoclast function at 10⁻⁶ M. These results suggest that taxanes have beneficial effects in the treatment of bone metastatic cancers. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Changes in bone turnover markers during 14-day 6° head-down bed rest

Osteoporosis caused by exposure to microgravity represents a serious clinical concern, but the mechanisms have yet to be fully elucidated. The present research aimed to elucidate the effects of microgravity environments on bone turnover, with a specific focus on changes in bone resorption markers such as type I collagen cross-linked N-telopeptides (NTx) and deoxypyridinoline (Dpyr), for which scant data are available regarding detailed time course. Methods using 6° head-down bed rest were utilized to simulate a microgravity environment. Eleven adult male volunteers underwent 6° head-down bed rest for 14 days; measurements were made of serum and urine Ca concentrations, in addition to osteocalcin (OC), bone alkaline phosphatase (ALP), NTx, and Dpyr as bone turnover markers. By the end of bed rest, concentrations of bone ALP had significantly increased, but OC displayed a tendency toward decrease. Concentrations of Dpyr significantly increased from day 6, remaining elevated until the end of bed rest. Concentrations of NTx significantly increased on day 13 and at the end of bed rest. Serum and urinary concentrations of Ca increased significantly at the end of bed rest. Bone ALP represents a relatively early marker of osteoblast differentiation at the matrix maturation phase and OC is a late marker in osteoblast differentiation at the calcification phase. The present results therefore suggest an absolute increase in bone resorption and normal or reduced bone formation, together causing prominent uncoupling and rapid bone loss after simulated microgravity. Moreover, the present results suggest that bone resorption is enhanced at an early stage of exposure to microgravity environments.     

Regulation of Osteoclasts by Calcitonin and Amphiphilic Calcitonin Conjugates: Role of Cytosolic Calcium

The peptide hormone calcitonin is a potent inhibitor of osteoclastic resorption, but it is unstable and poorly absorbed following oral administration. Conjugates of salmon calcitonin covalently linked to low-molecular-weight amphiphilic polymers show improved stability and absorption. The purpose of this study was to investigate the biological activity of these conjugates in vitro using rat osteoclasts and HEK-293 cells transfected with the C1a isoform of the calcitonin receptor. Salmon calcitonin or its conjugates (10 pM–10 nM) caused rapid arrest of osteoclast membrane ruffling and subsequent retraction. The same amphiphilic polymer attached to an unrelated protein had no effect on osteoclast morphology or motility. Since calcitonin-induced retraction of osteoclasts is thought to be mediated by Ca²⁺ signaling, we investigated the effects of calcitonin and its conjugates on cytosolic free Ca²⁺ concentration ([Ca²⁺]_i). In HEK-293 cells transfected with the calcitonin receptor, these agents induced transient elevations of [Ca²⁺]_i. However, the rise of [Ca²⁺]_i in HEK-293 cells occurred at concentrations 100–1000-fold higher than those required to elicit osteoclast retraction. To investigate the role of Ca²⁺ in osteoclast retraction, we preloaded cells with BAPTA to buffer changes in [Ca²⁺]_i. BAPTA decreased the initial rate of calcitonin-induced osteoclast retraction, but it did not affect the degree of retraction 2–3 hours following calcitonin, indicating that retraction is mediated primarily by Ca²⁺-independent processes. We conclude that calcitonin conjugates cause osteoclast retraction and [Ca²⁺]_i signaling in a manner similar to that elicited by calcitonin. Thus, orally bioavailable calcitonin conjugates show potential for use as antiresorptive agents.     

免疫细胞化学法检测乙型肝炎产妇分娩的死胎心脏组织乙型肝炎核心抗原的表达

目的 :观察乙型肝炎病毒 ( HBV)通过产妇传播在胎儿心脏组织中表达的情况。方法 :采集 40例乙型肝炎产妇分娩的死胎 ,常规尸检 ,取心脏组织 ,免疫细胞化学法检测乙型肝炎核心抗原 ( HBc Ag) ;回访孕妇产前静脉血 HBV的检测结果 ;采用百分率行 χ2检验。结果 :HBc Ag阳性颗粒在死胎的心内膜及心脏血管中呈点状及灶状分布 ,心肌细胞核不着色。乙型肝炎表面抗原 ( HBs Ag)、乙型肝炎 e抗原 ( HBe Ag)、乙型肝炎核心抗体( HBc Ab)皆为阳性的孕妇较 HBV单项阳性或 HBs Ag、乙型肝炎 e抗体 ( HBe Ab)、HB-c Ab皆为阳性的孕妇分娩的死胎心脏组织中 HBc Ag阳性率明显升高 ( P<0 .0 5) ;HBV单项阳性与 HBs Ag、HBe Ab、HBc Ab皆为阳性的孕妇分娩的死胎心脏组织中 HBc Ag阳性率比较 ,无显著差异 ( P>0 .0 5)。结论 :孕妇静脉血 HBs Ag、HBe Ag、HBc Ab皆为阳性是乙型肝炎病毒通过母婴垂直传播在胎儿心脏组织中表达的高危因素。     

运动调控骨代谢的关键膜上受体—G蛋白偶联受体48

G蛋白偶联受体48(GPR48)作为一富含亮氨酸重复序列的膜上七次跨膜受体,其LRR结构域与R-spondin1或Norrin结合形成复合体后可作用于下游关键因子可调控骨质疏松、阿尔茨海默病、高血压等疾病发生。骨作为重要的生理学和生物力学组织,由成骨细胞(OB)和破骨细胞(OC)分别主导的骨形成和骨吸收之间的拮抗及协同调控骨组织代谢平衡。骨细胞是力学刺激敏感细胞,调节应力加载后的骨适应性反应,而运动训练对骨产生的力学刺激可翻译成结构级联性生化反应(Wnt、c AMP/PKA/Atf4、OPG/RANKL/RANK等途径稳态表达),调控骨形成和/或骨吸收。并且,GPR48通过R-spondin1可直接调控以上信号途径的激活状态。那么,GPR48能否通过下游级联信号途径从而在运动影响骨代谢中起分子介导作用呢?探究GPR48在运动影响骨代谢中的作用及其分子介导机制,希望能进一步完善运动影响骨代谢的分子机制信号网络并为骨疾病诊治提供新靶点和非药物干预方式。     

骨质疏松症治疗药物研究进展

骨质疏松症是一种与增龄相关的代谢性骨病,主要是由成骨细胞介导的骨形成与破骨细胞介导的骨吸收之间的失衡所致.随着骨质疏松症患病率的逐年递增,如何防治骨质疏松已经成为我国重要公共健康问题.目前骨质疏松症治疗药物主要可分为促进骨形成剂和抑制骨吸收剂.本文主要就骨质疏松症治疗药物的研究进展作一综述,为临床治疗骨质疏松症提供理论...     

成人与青少年在内收上前牙的正畸治疗中牙根吸收的比较研究

目的：探讨成人与青少年在内收上前牙的正畸治疗中牙根吸收的差异。方法：选择运用直丝弓矫治技术矫治完成的上颌前突并拔牙矫治患者79例,其中成人患者38例,青少年患者41例。在其矫治前后的曲面断层片上对4个上颌切牙进行牙根形态观察及冠根长度的测量,以获得正畸治疗后牙根吸收的定性及定量资料。利用SPSS11.0软件对所得资料进行检验分析。结果：治疗后成人患者上颌中切牙的根吸收量大于侧切牙,成人患者上颌中切牙的根吸收量亦大于青少年患者,差异有统计学意义（P〈0.05）;同时,治疗后成人与青少年患者上颌中切牙的牙根吸收等级分布存在差异（P〈0.01）。结论：成人在上颌前牙的内收矫治中中切牙的根吸收风险较大。