甲状旁腺激素对成骨肉瘤细胞系SaOS-2细胞成骨活性的影响

目的 观察不同浓度重组甲状旁腺激素1～34(简称hPTH)对成骨肉瘤细胞系SaOS-2细胞(简称SaOS-2细胞)骨碱性磷酸酶(ALP)和骨钙素(BGP)mRNA表达的影响.方法 SaOS-2细胞正常传代培养,用0、1、10、100 nmol/L hPTH处理细胞,分别在12、24、48 h提取细胞总RNA,反转录合成cDNA,采用实时荧光定量PCR的方法测定ALP和BGP mRNA的表达.结果 ①hPTH的不同剂量、不同的作用时间以及二者的交互作用对ALP的表达量均有影响(F值分别为29.32、2.92、7.64,P均<0.05).干预48 h,0、1、10、100 nmol/L组ALP mRNA的表达量(0.78±0.43、0.71±0.05、0.75±0.19、0.76±0.14)均低于同浓度组的12、24 h(12 h:1.01±0.16、1.37±0.38、1149±0.16、2.52±0.70,24 h:1.80±0.47、1.30±0.36、1.27±0.17、1.17±0.11,P均<0.05).干预12 h,100 nmol/L组ALP mRNA的表达量高于0 nmol/L组(P<0.05);干预24 h,1、10、100nmol/L组SaOS-2细胞ALP mRNA的表达量均低于0 nmol/L组(P均<0.05).②hPTH的不同剂量、不同的作用时间以及二者的交互作用对SaOS-2细胞BGP mRNA的表达量均有影响(F值分别为8.26、10.33、5.51,P均<0.05).干预48 h,0、1、10、100 nmol/L组BGP mRNA的表达量(1.17±0.28、0.98±0.08、0.92±0.17、0.84±0.59)均低于同浓度组的12、24 h(12 h:1.01±0.14、1.21±0.18、1.34±0.30、1.68±0.62,24 h:1.71±0.35、1141±0.47、1.28±0.31、1.01±0.18,P均<0.05).干预12 h,100 nmol/L组BGP mRNA的表达量高于0、1nmol/L组(P均<0.05);干预24 h,10、100 nmol/L组SaOS-2细胞BGP mRNA的表达量均低于0 nmol/L组(P均<0.05),100 nmol/L组SaOS-2细胞BGP mRNA的表达量低于1 nmoL/L组(P<0.05);干预48 h,10、100 nmol/L组SaOS-2细胞BGP mRNA的表达量均低于0 nmol/L组(P均<0.05).结论 在体外培养条件下,hPTH在短时间作用下可以显著增强SaOS-2细胞的成骨活性,随着刺激时间的延长,可使成骨活性呈现下降的趋势.

Abstract：

Objective To observe the effects of recombinant human parathyroid hormone 1 to 34(referred to as hPTH) on the expression level of alkaline phosphatase(ALP) and bone gla protein(BCP) in human osteosarcoma cell line SaOS-2(referred to as SaOS-2 cells). Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different(F = 29.32, 2.92, 7.64, all P < 0.05). The expression level of ALP mRNA(0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14) of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h(1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P< 0.05) and 24 h (1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P< 0.05). The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control(P < 0.05); the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different (F = 8.26, 10.33, 5.51, all P< 0.05). The expression level of BGP mRNA(1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59) of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h( 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P< 0.05), and 24 h(1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH(all P< 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control(all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH(P < 0.05). Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.     

骨源性碱性磷酸酶与骨折愈合的相关性分析

目的探讨成人股骨干骨折患者血清中骨源性碱性磷酸酶（BALP）的活性与骨折愈合的关系。方法检测115例股骨干骨折术后第5、14天BALP水平,根据检测结果选择32例（高于平均值10%）为观察组,26例（低于平均值10%）为对照组。分别于术后1、3个月摄X线片观察两组骨折愈合情况。结果1个月时观察组骨痂量明显多于对照组,观察组骨折线较对照组模糊;3个月时观察组骨痂量亦多于对照组,观察组大部分患者骨折线消失或模糊不清,对照组大部分患者骨折线模糊,但未消失,少数患者骨折线仍清晰。结论BALP水平可判断骨折患者的骨形成能力和速度,可为骨折的病情监测、预后判断和治疗提供依据。     

血液透析影响血清碱性磷酸酶、骨硬化蛋白与骨代谢的机制

目的 研究血液透析影响血清碱性磷酸酶(ALP)、骨硬化蛋白与骨代谢的机制.方法 将163例受试者分为两组:血液透析组(血液透析患者,n=79),健康对照组(医院健康体检参与者,n=84);通过标准实验室方法测定血清和尿液中的肌酐水平;通过Western印迹检测骨硬化蛋白和血清ALP蛋白的表达;通过试剂盒检测骨代谢标志物...     

联合检测血清碱性磷酸酶和甲状旁腺激素对维持性血液透析患者全因及心血管事件死亡风险的预测价值

探讨联合检测血清碱性磷酸酶(ALP)和全段甲状旁腺激素(iPTH)水平对维持性血液透析(MHD)患者全因和心血管事件死亡风险的预测价值。入选MHD患者224例,检测患者血清iPTH、ALP等临床指标,随访2年,采用Cox回归分析全因死亡和心血管死亡的危险因素。结果显示,ALP水平与患者心血管事件死亡相关(HR=1.001, 95%CI 1.000～1.004,P=0.048),ALP≥106 IU/L是患者心血管事件死亡的独立危险因素(P<0.05)。联合检测ALP、iPTH与全因死亡相关(HR=1.002, 95%CI 1.000～1.004,P=0.033),若联合检测ALP-iPTH≥37.28是MHD患者全因死亡的独立危险因素(P<0.05)。血清iPTH水平与MHD患者全因死亡和心血管事件死亡无显著关系。     

中国恒河猴SHIVKU-1艾滋病模型中枢神经系统的病理研究北大核心CSCD

目的建立中国恒河猴SHIVKU-1艾滋病模型,观察感染SHIVKU-1的中国恒河猴中枢神经系统损伤的病理改变特点。方法用SHIVKU-1感染2只中国恒河猴(编号WSH01和WSH02),分别于感染后第72周和第70周出现严重艾滋病症状时执行安乐死。取脑组织的不同部位行常规病理切片,进行HE染色检查和免疫组织化学染色检查。结果 WSH01恒河猴感染晚期出现站立困难、行动迟缓、左半身控制能力丧失等明显的神经艾滋临床症状。WSH02恒河猴感染晚期全身耗竭,严重腹泻。HE染色检查WSH01恒河猴小脑、脑干、枕叶中可见多核巨细胞浸润和神经胶质小结形成;大脑皮层小胶质细胞增生;海马回神经元细胞减少、排列无序。病理诊断为艾滋病脑炎。WSH02猴仅在脑组织中观察到部分小胶质细胞结节,未见其他明显的神经病理损伤。免疫组化检查观察到在实验猴脑组织有两种主要的损伤形式,分别为典型的血管周病变和弥散性病变;大量活化巨噬细胞和小胶质细胞浸润和激活,多核巨细胞形成(CD68,CD163,Iba-1);损伤部位SHIVKU-1病毒蛋白P28强阳性(SIV p28);WSH01恒河猴海马回神经元细胞明显减少且排列无序(MAP2)。结论中国恒河猴SHIVKU-1感染导致中枢神经系统损伤,脑组织各部位表现出不同程度的病理改变,以大量巨噬细胞/小胶质细胞浸润、激活,并伴有病毒显著增殖为主。     

家兔去卵巢后血清酶和骨元素变化的相互关系

目的　研究去卵巢后家兔血清酶和骨元素的变化。方法　新西兰家兔１２ 只,７ 月龄。术前取血,测定血清碱性磷酸酶、酸性磷酸酶和骨特异性碱性磷酸酶的活性作为术前组。手术摘除二侧卵巢,饲养８ 个月后,取血清测定上述三种酶活性作为术后组。家兔处死,取髂骨作为实验组。另取１５ 个月龄完整家兔１２ 只,处死,取髂骨作为对照组,用中子活化方法测定髂骨的钙、镁、钠、锰、锶和钾元素的含量。结果　术前血清碱性磷酸酶、酸性磷酸酶和骨特异性碱性磷酸酶的活性分别为（２０ ．０６ ±８ ．２０）Ｕ／Ｌ、（１９．０６±５．４６）Ｕ／Ｌ和（１４．０４±１．６７）Ｕ／Ｌ,术后血清碱性磷酸酶、酸性磷酸酶和骨特异性碱性磷酸酶的活性分别为（１４．６０±４．６４）Ｕ／Ｌ、（１１ ．０４ ±４．１０）Ｕ／Ｌ和（４ ．４４±２ ．１０）Ｕ／Ｌ,摘除卵巢后上述各项生化指标均显著降低（ Ｐ＜ ０ ．００１）。家兔切除卵巢后实验组髂骨镁元素含量为（４．４４±０．１９）ｍｇ／ｇ,对照组髂骨镁元素含量为（５．４０±０ ．３２）ｍｇ／ｇ。实验组髂骨镁元素含量显著低于对照组（Ｐ＜ ０．００１） 。钙、钠、锰、锶和钾元素含量二组比较无显著差别。结论　成年家兔去卵巢８ 个月后,血清碱性磷酸酶     

骨特异性碱性磷酸酶联合Ⅰ型前胶原氨基端延长肽和其特殊序列在糖尿病骨质疏松中的应用

目前认为1型糖尿病可降低骨密度(BMD),导致骨质疏松(OP)〔1〕,但2型糖尿病对BMD的影响仍有争议〔2〕。本文拟探讨骨特异性碱性磷酶酶(BAP)、总Ⅰ型前胶原氨基端延长肽(TPINP)和其特殊序列(β-CTx)水平与2型糖尿病OP的关系。1材料与方法1.1一般资料按照1999年WHO糖尿病诊断标准,选择自我院内分泌科住院的2型糖患者60例(糖尿病组),年龄48～     

去卵巢大鼠骨形成参数和血清碱性磷酸酶的相关性研究

目的　观察去卵巢大鼠骨组织形态计量学参数和血清碱性磷酸酶 (ALP)之间的相关性。方法　 4个半月龄 SD大鼠双侧卵巢去除术后预防用药 90 d。用骨组织形态计量学方法测定大鼠胫骨组织近端松质骨形态计量学参数 ,并测定血清 ALP含量。结果　去卵巢组大鼠血清 ALP含量增加 ,骨形成参数增加 (P<0 .0 5)。去卵巢组和预防用药组骨形成参数与 ALP测量值之间有相关性 (P<0 .0 5)。去卵巢组的骨组织形态计量学参数与 ALP测量值之间的相关系数大于预防用药组。结论　去卵巢大鼠血清 ALP与骨形成参数之间存在相关性 ,这种相关性在给药后下降。     

快步行走对中老年女性骨密度、骨矿元素含量及碱性磷酸酶水平的影响

中老年女性受体内雌激素水平的影响,骨质疏松的患病率逐年增加,严重影响身体健康和生活质量.目前,越来越多的研究表明,适当的健身活动有助于延缓骨质疏松的发生、发展[1].但由于生活环境不同,很多健身活动不能广泛开展.快步行走对于场地没有要求,易于开展,已经逐渐成为最为百姓接受的健身活动.本研究拟通过检测长期进行快步行走的中老年女性体内骨矿元素钙(Ca)、磷(P)、镁(Mg)及血清碱性磷酸酶(ALP)水平,探讨快步行走对于防治中老年骨质疏松的作用及机制.     

β2肾上腺素能受体激动剂／拮抗剂对大鼠成骨细胞骨碱性磷酸酶比活性的影响

目的探讨β2肾上腺素能受体（ADRβ2R）激动N／拮抗剂对大鼠成骨细胞（0B）骨碱性磷酸酶（bALP）比活性的影响。方法取24h内新生sD大鼠的颅盖骨OB进行体外培养。将传至第3代的OB分成ADRβ2R激动剂组、ADRβ2R拮抗剂组和对照组,分别用不同浓度的ADRβ2R激动剂或ADRp：R拮抗剂对OB处理24、72和96h,检测bALP比活性的变化。结果ADRβ2R拮抗剂ICI-118551可以提高鼠OBbALP比活性;而ADRβ2R激动剂沙丁胺醇则降低鼠OBbALP比活性。两种调节作用均具有时间剂量依赖性。结论ADRβ2R激动N／拮抗剂可以通过调控OB骨形成指标bALP比活性变化,从而调节骨代谢。     

Verapamil increases serum alkaline phosphatase in hypertensive patients

In rats, verapamil decreases intestinal absorption of calcium, increases serum parathyroid hormone (PTH), and induces osteopenia. In this prospective study, verapamil 80-120 mg three times daily was given for 2 months to 20 patients with hypertension, and the effects on calcium homeostasis were recorded. This dose of verapamil significantly reduced supine systolic and diastolic blood pressure (+/- SD) from 158/100 +/- 9/8 mmHg to 146/89 +/- 14/8 mmHg (P = 0.001). Serum alkaline phosphatase (ALP) increased significantly from 2.77 +/- 1.06 mu kat l-1 to 3.19 +/- 1.22 mu kat l-1 (P = 0.004), and isoenzymes of ALP of skeletal origin appeared after verapamil treatment. The excretion of sodium in the urine increased (Na/creatinine ratio 8.95 +/- 6.01 before and 13.16 +/- 8.26 after verapamil; P = 0.04), while the excretion of calcium, phosphate and potassium was not changed. PTH was slightly increased at the end of verapamil treatment (1.09 +/- 0.54 vs. 0.98 +/- 0.74 microgram l-1; P = 0.07), and s-1,25(OH)2D3 was also somewhat increased (22.3 +/- 14.4 vs. 17.6 +/- 4.9 ng l-1; P = 0.26). Serum Ca was not affected by verapamil (before verapamil 2.43 +/- 0.11 mmol l-1, after verapamil 2.40 +/- 0.12 mmol l-1; P = 0.28). The increase in serum ALP demonstrates that verapamil affects bone cell metabolism in man. This effect could be secondary to the enhancement of PTH secretion.     

High frequencies of resting CD4+ T cells containing integrated viral DNA are found in rhesus macaques during acute lentivirus infections

We and others have reported that the vast majority of virus-producing CD4⁺ T cells during the acute infection of rhesus macaques with simian immunodeficiency virus (SIV) or CXCR4 (X4)-using simian/human immunodeficiency viruses (SHIVs) exhibited a nonactivated phenotype. These findings have been extended to show that resting CD4⁺ T lymphocytes collected from SIV- or X4-SHIV-infected animals during the first 10 days of infection continue to release virus ex vivo. Furthermore, we observed high frequencies of integrated viral DNA (up to 5.1 × 10⁴ DNA copies per 10⁵ cells) in circulating resting CD4⁺ T cells during the first 10 days of the infection. Integration of SIV DNA was detected only in memory CD4⁺ T cells and SHIVs preferentially integrated into resting naïve CD4⁺ T cells. Taken together, these results show that during the acute infection large numbers of resting CD4⁺ T cells carry integrated nonhuman primate lentiviral DNA and are the major source of progeny virions irrespective of coreceptor usage. Prompt and sustained interventions are therefore required to block the rapid systemic dissemination of virus and prevent an otherwise fatal clinical outcome.     

Abnormal bone turnover in monoclonal gammopathy of undetermined significance: analyses of type I collagen telopeptide,osteocalcin, bone-specific alkaline phosphatase and propeptides of type I and type III procollagens

Abstract: The main difference between monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM) is the presence of lytic bone destructions in the latter. About 20% of MGUS patients develop MM, and histomorphometric studies have shown disturbed bone turnover rates in some of these patients. This study was performed in order to evaluate whether serum analyses of the C-terminal telopeptide of type I collagen (ICTP), as a reflector of bone degradation, and of osteocalcin, bone-specific alkaline phosphatase (bAP) and the C-terminal propeptide of type I procollagen (PICP), as markers of bone formation, might give information on disturbancies of bone metabolism in MGUS. Furthermore, serum N-terminal propeptide of procollagen III (PIIINP) might give information on disturbances in collagen III metabolism in the bone marrow. In the 35 patients examined, serum ICTP was elevated in 12 patients (34%), serum PIIINP elevated in 6 patients (17%), serum osteocalcin elevated in 11 patients (31%), serum bAP elevated in 6 patients (17%), and serum PICP elevated in 4 patients (11%). Serum ICTP correlated significantly with PIIINP (r=0.72, p<0.001), and with serum osteocalcin (r=0.57, p<0.001) and serum bAP (r=0.51, p=0.002). These findings indicate disturbancies of bone turnover and affected collagen metabolism in some MGUS patients. Follow-up observation may reveal any prognostic value of these findings.     

我国恒河猴粪便腺病毒聚合酶基因片段的序列分析

目的研究非人灵长类动物猴子腺病毒的分子多样性。方法呆集圈养的恒河猴粪便样本，以腺病毒聚合酶基因为目的基因，用PCR3-法进行扩增，对PCR阳性产物进行克隆、测序，并进行系统进化分析。结果57只恒河猴粪便样本有12份存在腺病毒DNA，系统进化分析提示这些序列主要可分为两大组：类SAdV6组（2个非重复序列）和类SAdV一7组（9个非重复序列）。此外，有3个克隆，其最相似序列分别为：SAdV一1，SAdv_3和HAdV一52。结论研究证实腺病毒在非人灵长类粪便中普遍存在，揭示这些动物肠道内腺病毒的多样性以及系统进化情况。辇铺{司．帽河准．獬早限病毒：其闵相多样忡     

Influence of Naturally Occurring Simian Foamy Viruses (SFVs) on SIV Disease Progression in the Rhesus Macaque (Macaca mulatta) Model

We have investigated the influence of naturally occurring simian foamy viruses (SFVs) on simian immunodeficiency virus (SIV) infection and disease in Indian rhesus macaques. Animals were divided into two groups based upon presence or absence of SFV; in each group, eight monkeys were injected with SIV_mac239 virus obtained from a molecular clone and four were injected with medium. Blood was collected every two weeks for evaluation of SIV infection based upon T cell-subsets, plasma viral load, development and persistence of virus-specific antibodies, and clinical changes by physical examination and hematology. Comparative analysis of SFV+/SIV+ and SFV−/SIV+ monkey groups indicated statistically significant differences in the plasma viral load between 6–28 weeks, particularly after reaching plateau at 20–28 weeks, in the CD4⁺ and CD8⁺ T-cell numbers over the entire study period (2–43 weeks), and in the survival rates evaluated at 49 weeks. There was an increase in the plasma viral load, a decreasing trend in the CD4⁺ T cells, and a greater number of animal deaths in the SFV+/SIV+ group. The results, although based upon a small number of animals, indicated that pre-existing SFV infection can influence SIV infection and disease outcome in the rhesus macaque model. The study highlights consideration of the SFV status in evaluating results from SIV pathogenesis and vaccine challenge studies in monkeys and indicates the potential use of the SFV/SIV monkey model to study the dynamics of SFV and HIV-1 dual infections, recently reported in humans.     

Expression and kinetic changes of alkaline phosphatase and its isoenzymes in experimental rat hepatoma

AIM To explore the expression and changes of hepatoma specific alkaline phospatase (ALP) in rats during canceration.METHODS The ALPs and isoenzymes of rat livers and sera were investigated in SD hepatomas induced with 0.05% 2-fluoenylacetamide (2-FAA).RESULTS By pathological examination and biochemical analysis. ALPs were overexpressed in rat livers during canceration and then were secreted into blood. Serum total ALP activities, liver ALP specific activities (U/g) including soluble and membrane-combined ALP activities of each group were all significantly higher (P＜0.01) than those of control group. The average ratios of soluble ALP to membrane-combined ALP were increased significantly after 6 weeks. ALP isoenzymes of rat sera and livers showed 5 bands on PAGE: ALP-Ⅰ and ALP-Ⅱ were specific for normal liver and rat hepatoma tissues, the ALP-Ⅱ appeared in rat liver after 6 weeks and in sera after 8 weeks.CONCLUSIONS ALP with carcino-embryonic protein was overexpressed in hepatoma tissues; the abnormal ALP-Ⅱ of ALP isoenzymes in sera and liver of rats can be used as a tumor marker for early diagnosis of rat hepatoma.     

Quantitative changes in acid phosphatase,alkaline phosphatase and 5‘-nucleotidase activity in rat liver after experimentally induced cholestasis

ABSTRACT— Acid phosphatase, alkaline phosphatase and 5′-nucleotidase activities were analyzed cytophotometrically in cryostat sections of rat liver up to 8 weeks after ligation and transsection of the common bile duct. Ligation resulted in cholestasis and induced alterations in both localization and activity of the enzyme investigated. The cellular distribution but not the activity of acid phosphatase changed in liver parenchyma. In control liver, the final reaction product was localized as discrete granules in the bile canalicular region of hepatocytes. The final reaction product was precipitated more diffusely within the cytoplasm after induction of cholestasis, most probably due to increased fragility of lysosomal membranes. In control liver, alkaline phosphatase activity was low and localized in the bile canalicular plasma membranes only. The total parenchymal activity increased threefold after the induction of cholestasis and is considered to be a compensatory mechanism in order to enhance the excretion of bile salts from hepatocytes. 5′-Nucleotidase was present at the bile canalicular and sinusoidal surfaces of plasma membranes of hepatocytes in control liver; total activity in pericentral areas was significantly higher than in periportal areas. Induction of cholestasis resulted in higher total activity and redistribution of the activity over all three surfaces of the plasma membranes, whereas heterogeneity over the different zones of the acinus disappeared. The appearance of the enzyme at lateral plasma membranes is suggested to be related to the formation of new sites for bile salt transport out of the hepatocytes. With respect to all three enzymes studied, alterations of liver parenchymal cells due to a disturbed bile transport were already established during the first week of cholestasis.     

实验性牙周炎大鼠牙龈组织中碱性磷酸酶水平变化及意义

目的观察碱性磷酸酶（ALP）水平变化在牙周炎发病中的作用。方法选用40只健康SD大鼠随机分为对照组和观察组各20只,观察组建立牙周炎模型,对照组常规饲养;分别于造模术后4周和8周各处死20只,采用酶标仪测定牙龈组织中ALP水平,观察牙槽骨吸收情况及牙周组织学改变。结果造模术后4周和8周时观察组牙龈组织中ALP水平均明显高于对照组（P〈0．01）,且术后8周高于4周（P〈0．01）;观察组有明显牙槽骨吸收及牙周组织炎症细胞浸润,对照组则无。结论ALP水平变化可能在牙周炎发病过程中起重要作用。