Mechanism of Sclerostin/Lrp4 in the regulation of vascular smooth muscle cell calcification

Objective To investigate the possible mechanism of sclerostin/Lrp4 in calcification of VSMC induced by high phosphorus and the protective effect of Ginkgo biloba extract. Methods Aortic vascular smooth muscle cells (VSMCs) of SD rats were extracted and identified. VSMCs were divided into normal control group, high phosphorus induced calcification group (10 mmol/L β-glycerophosphate+50 μg/ml ascorbic acid), and high phosphorus induced calcification+Ginkgo biloba extract intervention group (10 mmol/L β-glycerophosphate+50 μg/ml ascorbic acid+0.5 mg/ml GBE), cultured in different mediums for 14 days. Vonkossa staining and alizarin red staining were used to detect the calcification of VSMCs. The mRNA level of BGP was detected by real time PCR, and the protein expressions of sclerostin and Lrp4 were detected by Western blot. Results Compared with normal control group, vonkossa staining and alizarin red staining showed significant calcium deposition in calcification group. Compared with calcification group, calcium salt deposition was significantly reduced in GBE treatment group. Real time PCR results showed β-catenin and BGP mRNA expressions in VSMC calcification group were higher than those in normal control group (P＜0.05). mRNA expressions of β-catenin and BGP in GBE treatment group were lower than those in calcification group (all P＜0.05). Compared with normal control group, the protein expression of sclerostin was increased, but the protein expression of Lrp4 was decreased in calcified group (all P＜0.05). Compared with calcification group, the protein expression of sclerostin decreased and the protein expression of Lrp4 increased in GBE treatment group (all P＜0.05). Conclusions High phosphorus can induce VSMC calcification by activating Wn/β-catenin signaling pathway. Sclerostin/Lrp4 is involved in hyperphosphine-induced VSMC calcification. GBE can reduce the high phosphorus induced VSMC calcification by regulating the Wnt/β-catenin signaling pathway.     

Pregnancy in patients on maintenance hemodialysis: seven cases report and 169 reported cases review

Objective To analyze and summarize the cases of pregnancy on maintenance hemodialysis (MHD), and review the literature. Methods Seven cases of pregnancy and childbirth in patients on MHD in the First Affiliated Hospital of Zhejiang University from Jan 2009 to Dec 2017 were analyzed, and the literature about pregnancy in patients on MHD reported in Pubmed and Web of Science database was retrieved. Both maternal and fetal outcome were studied. Results There are seven pregnant MHD patients in this center, among whom six patients went through a smooth pregnancy and one patient had intra-uterine fetal death at 14 weeks of pregnancy. The six patients had preterm labor. Among six fetuses, four grew in good health and developed well, one had physical development retardation and one had heart malformation at born. In the literature, 169 cases reported pregnant patients and 182 fetuses were evaluated, of which 145 live infants were delivered, 79.67% overall fetal survival rate, with gestational age of (32.94±3.34) weeks. In 164 delivered fetuses, 147 were preterm labor (89.63%). The weight of live births was (1824±609) g. There were no maternal deaths. Fetal survival rate was zero in＜20 weeks of gestational age, 20-24 weeks was 14.29%, 25-27 weeks was 45.45%, ≥28 weeks was 94.44%. The fetal survival rate was higher in pregnant woman receiving hemodialysis≥28 hours weekly compared to women receiving ＜16 hours weekly (92.31% vs 52.94%, P=0.02). Conclusions There are still a very high maternal and fetal complication rate in hemodialysis patients, especially in fetus. Gestational age≥28 weeks has a high fetal survival rate. Intensive dialysis during pregnancy may benefit higher fetal survival rate.     

Differential expression of microRNAs related to apoptosis in human osteoblasts induced by sodium fluoride北大核心

BACKGROUND: Long-term excessive intake of fluoride, especially through drinking water, can cause chronic fluorosis of bone. The disease can lead to bone damage and deformity, and is difficult to recover. Unfortunately, we have not developed a noninvasive or minimally invasive method for its early diagnosis. OBJECTIVE: To observe the expression of apoptosis-related miRNAs under the action of excessive fluorine in human osteoblasts. METHODS: The fluorine model was established in the human osteoblasts by cultured with 20 and 40 mg/L sodium fluoride for 24 and 48 hours, respectively. The expression levels of apoptosis-related miRNAs were determined by PCR array. RESULTS AND CONCLUSION: After 24-hour treatment of sodium fluoride, 48 kinds of miRNAs were upregulated and 4 ones were down-regulated in the osteoblasts. After 48-hour treatment of sodium fluoride, 21 kinds of miRNAs were upregulated and 2 ones were down-regulated. It showed that nine up-regulated miRNAs and one down-regulated miRNA were same in two periods. The 10 miRNAs are selected for target gene analysis on bioinformatics software that refer to the effect of anti-apoptosis and pro-apoptosis, which is of great significance for the early identification of skeletal fluorosis. © 2018, Journal of Clinical Rehabilitative Tissue Engineering Research. All rights reserved.     

Lentiviral Vectors and Adeno‐Associated Virus Vectors: Useful Tools for Gene Transfer in Pain Research

Pain, especially chronic pain, has always been a heated point in both basic and clinical researches since it puts heavy burdens on both individuals and the whole society. A better understanding of the role of biological molecules and various ionic channels involved in pain can shed light on the mechanism under pain and advocate the development of pain management. Using viral vectors to transfer specific genes at targeted sites is a promising method for both research and clinical applications. Lentiviral vectors and adeno‐associated virus (AAV) vectors which allow stable and long‐term expression of transgene in non‐dividing cells are widely applied in pain research. In this review, we thoroughly outline the structure, category, advantages and disadvantages and the delivery methods of lentiviral and AAV vectors. The methods through which lentiviral and AAV vectors are delivered to targeted sites are closely related with the sites, level and period of transgene expression. Focus is placed on the various delivery methods applied to deliver vectors to spinal cord and dorsal root ganglion both of which play important roles in primary nociception. Our goal is to provide insight into the features of these two viral vectors and which administration approach can be chosen for different pain researches. Anat Rec, 301:825–836, 2018. © 2017 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.     

The Effects of Calcium Supplementation on Verified Coronary Heart Disease Hospitalization and Death in Postmenopausal Women: A Collaborative Meta‐Analysis of Randomized Controlled Trials

Calcium supplementation, particularly with vitamin D, has been an approved public health intervention to reduce fracture risk. Enthusiasm for this intervention has been mitigated by meta‐analyses suggesting that calcium supplementation with or without vitamin D increases myocardial infarction (MI) risk; however, concern has been raised over the design of these meta‐analyses. We, therefore, undertook a meta‐analysis of randomized controlled trials with placebo or no‐treatment control groups to determine if these supplements increase all‐cause mortality and coronary heart disease (CHD) risk including MI, angina pectoris and acute coronary syndrome, and chronic CHD verified by clinical review, hospital record, or death certificate in elderly women. The Cochrane Central Register of Controlled Trials, MEDLINE, and EMBASE databases were searched from January 1, 1966, to May 24, 2013, for potentially eligible studies, reference lists were checked, and trial investigators were contacted where additional unpublished data were required. The search yielded 661 potentially eligible reports of which 18 met the inclusion criteria and contributed information on 63,563 participants with 3390 CHD events and 4157 deaths. Two authors extracted the data independently with trial data combined using random‐effects meta‐analysis to calculate the relative risk (RR). Five trials contributed CHD events with pooled relative RR of 1.02 (95% confidence interval [CI], 0.96–1.09; p = 0.51). Seventeen trials contributed all‐cause mortality data with pooled RR of 0.96 (95% CI, 0.91–1.02; p = 0.18). Heterogeneity among the trials was low for both primary outcomes (I² = 0%). For secondary outcomes, the RR for MI was 1.08 (95% CI, 0.92–1.26; p = 0.32), angina pectoris and acute coronary syndrome 1.09 (95% CI, 0.95–1.24; p = 0.22) and chronic CHD 0.92 (95% CI, 0.73–1.15; p = 0.46). In conclusion, current evidence does not support the hypothesis that calcium supplementation with or without vitamin D increases coronary heart disease or all‐cause mortality risk in elderly women. © 2014 American Society for Bone and Mineral Research.     

胫骨残端长度对前交叉韧带重建术后本体觉恢复的影响

目的:研究保留韧带胫骨残端的长度对前交叉韧带重建术后膝关节本体觉恢复的影响.方法:自2011年3月至2011年12月于我院骨科就诊的42例前交叉韧带重建患者分为3组,每组14例.A组保留韧带胫骨残端长度15～20 mm,男8例,女6例,平均年龄(28.2±6.6)岁;B组保留韧带胫骨残端5～10 mm,男9例,女5例,平均年龄(27.9±6.4)岁;C组保留韧带胫骨残端小于2 mm,男9例,女5例,平均年龄(28.6±6.8)岁.治疗前后采用Lachman试验、前抽屉试验对稳定性进行评价,行Lysholm评分和Tegner评分评价关节功能,采用非负重位被动重复位置法对膝关节本体觉进行检测.结果:3组患者术后6个月Lachman试验、前抽屉试验均为阴性,12、18个月3组各1例前抽屉试验呈弱阳性.术后18个月Lysholm和Tegner评分均高于术前,而术后18个月3组间差异无统计学意义.屈曲20°和伸直80°、20°测试重复位置觉显示,3组在治疗前后差异无统计学意义.屈曲50°和伸直50°测试重复位置觉显示,A组和B组在术后6个月较治疗前好转,且明显优于C组;术后18个月3组差异无统计学意义.屈曲80°测试重复位置觉显示,A组和B组在术后12个月较治疗前好转,且明显优于C组;术后18个月3组差异无统计学意义.结论:保留韧带胫骨残端有助于前交叉韧带重建术后早期本体觉的恢复,残端长度以5～10 mm为宜.     

Cervical Vagus Nerve Stimulation Improves Neurologic Outcome After Cardiac Arrest in Mice by Attenuating Oxidative Stress and Excessive Autophagy

BackgroundCerebral ischemia and reperfusion (I/R) induces oxidative stress and activates autophagy, leading to brain injury and neurologic deficits. Cervical vagus nerve stimulation (VNS) increases cerebral blood flow (CBF). In this study, we investigate the effect of VNS-induced CBF increase on neurologic outcomes after cardiac arrest (CA).Materials and MethodsA total of 40 male C57Bl/6 mice were subjected to ten minutes of asphyxia CA and randomized to vagus nerve isolation (VNI) or VNS treatment group. Eight mice received sham surgery and VNI. Immediately after resuscitation, 20 minutes of electrical stimulation (1 mA, 1 ms, and 10 Hz) was started in the VNS group. Electrocardiogram, blood pressure, and CBF were monitored. Neurologic and histologic outcomes were evaluated at 72 hours. Oxidative stress and autophagy were assessed at 3 hours and 24 hours after CA.ResultsBaseline characteristics were not different among groups. VNS mice had better behavioral performance (ie, open field, rotarod, and neurologic score) and less neuronal death (p < 0.05, vs VNI) in the hippocampus. CBF was significantly increased in VNS-treated mice at 20 minutes after return of spontaneous circulation (ROSC) (p < 0.05). Furthermore, levels of 8-hydroxy-2?-deoxyguanosine in the blood and autophagy-related proteins (ie, LC-3Ⅱ/Ⅰ, Beclin-1, and p62) in the brain were significantly decreased in VNS mice. Aconitase activity was also reduced, and the p-mTOR/mTOR ratio was increased in VNS mice.ConclusionsOxidative stress induced by global brain I/R following CA/ROSC leads to early excessive autophagy and impaired autophagic flux. VNS promoted CBF recovery, ameliorating these changes. Neurologic and histologic outcomes were also improved.