维生素D受体在1，25（OH）2D3调节人骨肉瘤细胞系HOS—8603增殖中的作用

目的：研究维生素D3受体（VDR）在1,25（OH）2D3及其类似物调节人骨肉瘤细胞系HOS-8603增殖中的作用。方法：用RT-PCR和免疫组织化学技术分别在mRNA和蛋白水平上明确HOS-8603细胞中是否有VDR表达;瞬时转染VDR报告基因技术观察HOS-8603细胞中VDR的功能活性;并进一步利用稳定表达VDR反义mRNA的细胞株VDRas3细胞研究VDR被阻断后细胞增殖以及基因转录的变化。结果：HOS-8603细胞有VDR表达,此内源性VDR具有激素依赖性的转录激活活性。在内源性VDR被阻断后,1,25（OH）2D3及其类似物对细胞增殖的抑制作用以及对VDR的靶基因p21 mRNA表达的诱导作用均明显减弱。结论：1,25（OH）2D3及其类似物对HOS-8603细胞增殖抑制作用是由VDR介导的。     

改进的人血清1,25（OH）2D3的放射受体测定法

改进的人血清１，２５（ＯＨ）２Ｄ３的放射受体测定法刘怀成周学赢孟迅吾邢小平（中国医学科学院北京协和医院内分泌科，中国协和医科大学内分泌研究中心，北京１００７３０）我们于１９８９年在国内首先报道了不需高压液相层析（ＨＰＬＣ）的１，２５（ＯＨ）２Ｄ３的测...     

维生素D3的免疫调节作用

维生素D3及其衍生物是维系体内钙磷平衡最重要的调节因子,近期随着研究的深入,已逐渐发现除上述功能外,它还具有重要的免疫调节作用,在免疫细胞的转化、信号的转导等方面均发现其存在独特的功能,对其免疫学功能方面的探讨已成为研究的热点之一.     

原发性肝癌患者血清25-羟基维生素D3水平与免疫抑制因子IL-10的关系

目的 探究原发性肝癌患者血清25-羟基维生素D3[25-（OH）D3]的变化及其与免疫抑制因子IL-10的关系。方法 选取2014年2月~2019年7月于我院就诊的60例原发性肝癌患者作为疾病组,另外选择同期60名健康体检人群作为健康对照组,采用电化学发光法测定两组血清25-（OH）D3浓度,酶联免疫吸附试验测定血清IL-10浓度。Spearman法分析25-（OH）D3与IL-10间的相关性。结果 疾病组血清25-（OH）D3浓度低于健康对照组[（13.65±7.92）ng/ml vs （26.15±8.33）ng/ml],差异有统计学意义（P＜0.05）;疾病组IL-10浓度高于健康对照组[（89.71±26.59）ng/ml vs （26.18±8.15）ng/ml],差异有统计学意义（P＜0.05）。血清25-（OH）D3可能与淋巴结转移和肿瘤分期密切相关,与IL-10浓度呈负相关（r=-0.568,P＜0.05）。结论 原发性肝癌患者存在25-（OH）D3缺乏现象,且与肿瘤进展和免疫抑制密切相关。     

维生素D3受体对hsp90β基因表达的调控作用

鉴于热休克蛋白９０β（ｈｓｐ９０β）基因内含子中含有维生素Ｄ３受体（ＶＤＲ）结合位点，为探讨作为核受体家族成员的ＶＤＲ是否对核受体特异分子伴侣的ｈｓｐ９０β基因的表达具有调控作用，我们开展了本项研究。分别将野生型ＶＤＲ、含Ｎ端（１～１３３氨基酸残基）及Ｃ端（２８１～４２７氨基酸残基）片段的ＶＤＲ突变体真核表达质粒与人ｈｓｐ９０β基因调控片段（－１０３９／＋１５３１）介导的氯霉素乙酰基转移酶（ＣＡＴ）报告基因质粒共转染Ｊｕｒｋａｔ细胞，检测正常及经热休克（４２℃，１ｈ）处理后细胞裂解液中ＣＡＴ活性。结果表明ＶＤＲＮ端增强、而Ｃ端抑制ｈｓｐ９０β的组成性表达；在热诱导条件下野生型ＶＤＲ对ｈｓｐ９０β的表达有一定的抑制作用，而其Ｃ端片段的抑制较强。为进一步研究ＶＤＲ对细胞内源性热休克基因表达的影响，我们用ＲＴＰＣＲ方法研究了ＶＤＲ的对细胞内ｈｓｐ９０β基因ｍＲＮＡ水平的影响，发现ＶＤＲ过表达对ｈｓｐ９０β的热诱导表达明显抑制。结果提示ＶＤＲ对ｈｓｐ９０β基因的组成性和热诱导表达的调控机制不同。     

维生素D3受体在人胎脑表达的研究

为研究维生素D3受体(VDR)在正常人胎脑的分布,进而探讨1,25-二羟维生素D3[1,25-(OH)2D3]与正常人胎脑发育的关系,本实验按合法程序收集2～7月龄正常人胎脑,用免疫组织化学方法研究VDR在正常人胎脑的时空表达情况。结果显示:阳性产物在海马结构存在于胞核,3月龄出现表达的高峰,此后逐渐下降;在额叶皮质以及中脑部位的胞核和胞浆都可存在,其表达随胎龄增长。以上结果表明VDR在2～7月龄的人胎脑额叶皮质、海马结构和中脑有表达,且各部位的表达模式不同,提示1,25-(OH)2D3通过VDR可在人胎脑不同部位发育中发挥作用。     

维生素D通过下调谷氨酰胺合成酶抑制乳腺癌细胞增殖

目的 探究维生素D对非三阴性乳腺癌(non-triple negative breast cancer, Non-TNBC)和TNBC癌细胞增殖的影响及分子机制。方法 收集TNBC和Non-TNBC患者乳腺组织,原代培养TNBC和Non-TNBC患者乳腺细胞;免疫荧光检测乳腺细胞雌激素受体(ER)、孕激素受体(PR)和HER2蛋白表达;免疫荧光检测Non-TNBC和TNBC乳腺癌组织和乳腺细胞中VDR蛋白表达;CCK-8检测细胞活力变化;流式细胞术检测细胞增殖和细胞周期的变化;光学比色法检测细胞谷氨酰胺合成酶(glutamine synthetase, GS)活力的变化;ELISA检测细胞培养上清和胞内谷氨酰胺的水平;Western blot检测细胞GS和VDR蛋白表达的变化;CHIP-PCR检测VDR对GS的转录调控。结果 TNBC患者外周血中维生素D水平和癌组织VDR蛋白表达明显低于Non-TNBC患者(P<0.05)。相比于原代Non-TNBC乳腺癌细胞,原代TNBC患者乳腺癌细胞低表达雌激素受体(ER)、孕激素受体(PR)和HER2蛋白表达;TNBC乳腺癌细胞VDR表达水...     

原发ITP患者外周血维生素D3水平与维生素D受体表达的相关性研究

目的 研究成人原发免疫性血小板减少症(ITP)患者血清中25-羟维生素D3[25-(OH)-D3]、维生素D活性指标与维生素D受体表达的相关性.方法 选择开滦总医院2017年1月至2019年12月收治的30例ITP患者作为研究对象,设为ITP组;选择同期来院体检的30例健康体检者作为对照组,抽取外周血,检测血清中25-...     

3D可视化联合3D打印在肝癌大部分肝切除术中的应用

目的:探讨基于3D重建系统软件的肝体积评估和3D可视化、3D打印辅助肝癌大部分肝切除术的应用价值。方法:将符合要求的肝癌行大部分肝切除术患者46例,随机分为观察组和对照组,每组23例。观察组（3D组）患者采用3D可视化技术和3D打印模型进行围手术期规划和指导,主要基于肝体积评估等术前规划和3D可视化分析、3D打印指导肝切除术手术;对照组（CT组）患者采用传统CT资料进行肝体积评估等术前规划、CT二维影像资料指导肝切除术。观察指标:虚拟切除肝体积、实际切除肝体积、残肝体积、标准残肝体积比、手术时间、术中出血量、术后并发症、患者满意度等。结果:3D组与CT组虚拟切除肝体积与实际切除肝体积、虚拟（术前）残肝体积与实际（术后）残肝体积比较,差异均无统计学意义（P>0.05）,相关性分析显示虚拟切除肝体积与实际切除肝体积呈正相关性（3D组r=0.990, P<0.001;CT组r=0.943, P<0.001）。3D组与CT组虚拟残肝体积比、实际残肝体积比比较,差异均无统计学意义（P>0.05）,且相关性分析显示呈正相关性（3D组r=0.931, P<0.001;CT组r=0.902, P<0.001)。3D组术中出血量少于CT组（P<0.05）,3D组患者满意度优于CT组（P<0.05）。两组患者手术时间、术后并发症等比较,差异无统计学意义（P>0.05）。结论:3D重建系统软件和CT软件在评估肝癌大部分肝切除术的肝体积均可行、准确,具有很好的临床应用价值,有助于肝切除术的安全实施。3D可视化联合3D打印在围手术规划可减少手术出血,提高患者满意度,在临床应用中具有潜在优势。     

维生素D在哮喘发病机制中的作用

生素D除了参与钙磷代谢和调节骨骼稳态作用外,还具有调节机体免疫等广泛的生物学调节功能。维生素D不足使免疫细胞增殖和分化偏移而产生异常免疫反应。哮喘是由多种细胞（包括肥大细胞、嗜酸性粒细胞和T淋巴细胞）参与的慢性气道炎症,也是一种由遗传和环境因素共同作用的复杂多基因遗传病。近些年研究表明,维生素D缺乏会增加哮喘易感性和严重程度,与哮喘的遗传和非遗传因素共同参与哮喘的发病。     

活性维生素D3联合力学拉伸对成骨细胞MC3T3-E1增殖、分化及OPG和RANKL表达的影响

目的体外模拟成骨细胞在体内的生存环境,考察活性维生素D3(VD3)、力学拉伸以及两者联合对成骨细胞MC3T3-E1增殖、分化及破骨细胞抑制因子(OPG)和破骨细胞分化因子(RANKL)表达的影响。方法将10 nmol/L VD3、间断性力学拉伸以及两者联合作用于成骨细胞。流式细胞术检测细胞增殖。荧光探针试剂盒检测碱性磷酸酶(ALP)活性。实时定量PCR检测核心转录因子Runx2、OPG、RANKL mRNA水平,Western blotting检测其蛋白表达。结果 VD3抑制成骨细胞增殖,力学拉伸不能改变这种抑制效应。力学拉伸和VD3单独作用成骨细胞均能增加ALP活性及提高ALP、Runx2 mRNA水平,但当联合刺激后这些指标均降低,且成强度依赖性。力学拉伸增加OPG/RANKL比值,增强成骨作用,联合VD3后,OPG/RANKL比值下降。结论力学拉伸能有效诱导成骨分化,增加骨形成。VD3与力学拉伸联合抑制成骨细胞增殖和分化,并通过增加RANKL表达而影响骨重建。研究结果为骨质疏松及相关骨疾病的理论和治疗提供有意义的探索。     

Effects of vitamin D on the growth of normal and malignant B-cell progenitors.

As the effects of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25-(OH)2-D3) (VD, calcitriol) on the proliferation and differentiation potential of normal and leukaemic cells in vitro of myeloid lineage are known, we investigated the response to VD on the growth of both normal and malignant lymphoid progenitors. Effects of vitamin D on normal human lymphoid progenitors and B lineage acute lymphoblastic leukaemia (ALL) progenitors were assessed by using an in vitro cell colony assay specific for either B or T cell lineages. The expression of VDR on B untreated malignant progenitors at diagnosis was investigated by RT-PCR analysis. VD induced a significant inhibition of normal lymphoid cell progenitors growth of both T and B lineage. VD inhibited significantly also the growth of malignant B cell lineage lymphoid progenitors, without inducing cytotoxic effect. As it has been reported that VD effects on activated lymphocytes are mediated by 1,25-(OH)2-D3 nuclear receptor (VDR), we investigated VDR expression on malignant B cell progenitors. We did not detect VDR expression on these cells examined at diagnosis. We demonstrated that VD inhibited in vitro the clonogenic growth of both normal and malignant lymphoid B cell progenitors and that this inhibitory effect on malignant B cell progenitors was not related to VDR. Our work contributes to understanding of the mechanism of action of this hormone in promoting cellular inhibition of clonal growth of malignant lymphoid B cell progenitors, suggesting that the regulation of some critical growth and differentiation factor receptors could be a key physiological role of this hormone.     

1alpha,25-dihydroxyvitamin D3 increases IgA serum antibody responses and IgA antibody-secreting cell numbers in the Peyer's patches of pigs after intramuscular immunization

Pigs were injected intramuscularly (i.m.) twice with human serum albumin (HSA) with or without 1alpha,25-dihydroxyvitamin D3[1alpha,25(OH)2D3] with a 5-week interval. The supplementation of 1alpha,25(OH)2D3 enhanced the HSA-specific IgA serum antibody response but decreased the IgM, IgG, IgG1 and IgG2 responses. Furthermore, higher numbers of HSA-specific IgA antibody-secreting cells were obtained in systemic lymphoid tissues (local draining lymph node, spleen and bone marrow) as well as in Peyer's patches and lamina propria of the gut (GALT). In addition, the in vivo mRNA expression for Th1 [interferon (IFN)-gamma, interleukin (IL-2)], Th2 (IL-4, IL-6 and IL-10) and Th3 [transforming growth factor (TGF)-beta] cytokines as well as the percentage of different cell subsets (CD2+, CD4+, CD8+, IgM+, MHC II+, CD25+) of monomorphonuclear cells from the local draining lymph node were determined at different time-points after the i.m. immunizations. Cytokine profiles did not resemble a typical Th-cytokine profile using 1alpha,25(OH)2D3: higher levels of IL-10 and significantly lower levels of IL-2 were observed the first day after the primary immunization. However, significantly higher levels of IL-2 and significantly lower levels of IFN-gamma were observed the first day after the second immunization. Furthermore, after the second immunization TGF-beta mRNA expression decreased more quickly in the 1alpha,25(OH)2D3 group. This difference became significant 7 days after the second immunization. One week later a significantly higher percentage of CD25+ cells was observed in this group, indicating more activated T and B cells using the steroid hormone. These results suggest that in pigs the addition of 1alpha,25(OH)2D3 to an intramuscularly injected antigen can enhance the antigen-specific IgA-response and prime GALT tissues, but the relation with cytokines and cell phenotype in the local draining lymph node needs further clarification.     

1, 25二羟基胆钙化醇诱导大鼠骨髓单核细胞向破骨细胞转化的机制

目的:探讨1,25二羟基胆钙化醇(1,25(OH)2D3)诱导大鼠骨髓单核细胞向破骨细胞转化时明胶酶表达及其参与骨陷窝形成机制。方法:分离乳鼠骨髓内细胞,诱导生成破骨样细胞。姬姆莎、抗酒石酸酸性磷酸酶(TRAP)染色鉴定。扫描电镜观察诱导出的细胞贴附于骨片上形成的骨陷窝,明胶酶谱检测细胞培养液中明胶酶表达水平。结果:单核细胞经1,25(OH)2D3诱导,第9日生成大量的破骨样细胞。姬姆莎染色显示出多核(≥3个),TRAP染色阳性,扫描电镜观察破骨细胞在骨片上培养时产生骨陷窝。1,25(OH)2D3组基质金属蛋白酶2(MMP-2)表达水平增加显著。结论:1,25(OH)2D3诱导单核细胞产生大量破骨细胞。参与骨陷窝形成的明胶酶是MMP-2。这可能是破骨细胞通过某些机制,促进其他非破骨细胞分泌有活性的MMP-2参与噬骨。     

25(OH)D3与糖尿病视网膜病变的关系研究

目的 探讨25(OH)D3与糖尿病视网膜病变之间的关系。方法 回顾性分析2016年1月~2017年3月我科收治的2型糖尿病患者83例,根据患者是否合并视网膜病变,分为观察组43例和对照组40例。所有患者检测并比较血糖（FBG）水平、总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白（HDLC）、低密度脂蛋白（LDL-C）、糖化血红蛋白（HbA1c）以及25(OH)D3的水平。结果 两组患者在TG（t=5.572,P=0.037）、HDLC（t=5.548,P=0.037）、HbA1c（t=6.627,P=0.012）、25（OH）D3（t=9.738,P=0.000）等临床生化资料比较中差异均具有统计学意义（P＜0.05）。对25(OH)D3的Pearson进行相关性分析后发现,25(OH)D3与T2DM病程（r=0.163,P=0.027）、TC（r=0.170,P=0.025）、HDLC（r=0.177,P=0.023）、LDLC（r=0.185,P=0.015）具有高度相关性;偏相关显示,25(OH)D3与TC、HDLC、LDLC具有相关性。结论 25(OH)D3水平能够反映糖尿病视网膜病变患者的疾病发展状态,在诊断及评估中具有一定的参考价值。     

Intravenous 1α, 25[OH]2 vitamin D3 (calcitriol) pulse therapy for bone lesions in a murine model of chronic cadmium toxicosis

The aim of the present study was to clarify the therapeutic effects of 1alpha, 25[OH]2 vitamin D3 (calcitriol) pulse injection on bone lesions induced in a rat model of chronic cadmium toxicosis. Ovariectomized (OVX) and control-operated (sham-OVX) rats were given repeated intravenous injections of 0.5 mg/kg/day CdCl2 for 70 weeks. The rats were then treated intravenously with 0.02 microg/kg/day calcitriol 3 days per week for 8 weeks. CdCl2 treatment induced increases in osteoid volumes of the femur cortex and trabecula. This change was accompanied by an increase in the volume of iron deposition at the mineralization front of the trabeculae and a reduction in mineral density. Abnormalities of bone metabolic parameters, which were increases in the blood calcium, inorganic phosphorous, bone-specific alkaline phosphatase, parathyroid hormone (PTH) and osteocalcin levels, and in the urine deoxypyridinoline (D-PYR) level, were also induced. Calcitriol treatment increased the blood calcium and inorganic phosphorous levels, and reduced the blood PTH level. Decreases in blood tartrate-resistant acid phosphatase and urine d-PYR levels were also induced indicating that bone resorption was suppressed. The findings indicated that the increased osteoid volume of the cortex and Fe-deposition volume of the trabecula were improved. These effects or improvements were observed in the sham-OVX rats but not in the OVX rats.     

Inhibition of the in vitro growth of Plasmodium falciparum by D vitamins and vitamin D-3 derivatives

D vitamins are effective inhibitors of the in vitro intraerythrocytic growth of Plasmodium falciparum. Disappearance of the parasitemia was observed after 48 h contact between infected cells and 5 x 10(-6) M 1 alpha-hydroxycholecalciferol, 5 x 10(-5) M 25-hydroxycholecalciferol (25-OH-D-3), 1 alpha, 25-dihydroxycholecalciferol or 2.5 x 10(-4) vitamin D-2 and D-3. A 48 h pretreatment of healthy erythrocytes with 5 x 10(-5) M 25-OH-D-3 did not change their susceptibility to invasion by the parasite and their ability to support the growth of P. falciparum. Ionomycin, a calcium ionophore, and EGTA prevented parasite development at concentrations greater than 2 x 10(-7) M and 4 x 10(-4) M, respectively, but did not antagonize the inhibitory activity of 25-OH-D-3. Addition of 25-OH-D-3 for 12 or 24 h duration to synchronized cultures, showed that the drug had a schizonticidal action, but was without effect when parasites were in the ring form.     

河南省1～7岁健康儿童血清25(OH)D水平分析

目的 了解河南省1～7岁健康儿童维生素D营养状况,为儿童合理补充维生素D、防治维生素D缺乏提供科学依据.方法 采用分层整群随机抽样法,选取河南省六个地区的妇幼保健院和社区卫生服务站进行体检的1～7岁健康儿童392名,采用化学发光法检测血清25 (OH)D水平,比较不同年龄段、不同性别儿童维生素D不足与缺乏情况.结果 392名1～7岁健康儿童血清25(OH)D平均水平为42.7±15.8nmol/L,不足与缺乏率达59.4％ (233/392),男女童之间差异无统计学意义(=0.500,P=0.618;x2 =0.051,P=0.822);不同年龄组维生素D平均水平及其不足与缺乏比例差异具有统计学意义(F=12.748,P＜0.001;x2=37.077,P＜0.001);维生素D水平随年龄增长而降低,不足与缺乏比例随年龄增长而升高.结论 河南省1～7岁健康儿童维生素D营养状况较差,且与年龄密切相关,应适当增加儿童日照时间,加强合理补充维生素D的科普宣传.