人白介素24腺病毒载体的构建与鉴定

目的构建人白介素24（hIL-24）的腺病毒载体，获得hIL-24重组腺病毒子，为hIL-24进行肿瘤的基因治疗奠定基础。方法以pcDNA3．0-hIL-24重组质料为模板，PCR扩增hIL-24，酶切连接到带有GFP标记的pAdTrack—CMVR质粒上，PmeI线性化重组质粒pAdTrack—CMV-hIL-24，与腺病毒质粒pAdEasy-1共转化BJ5183细菌，获得重组腺病毒载体pAdEasy-1-pAdTrack-CMV-hIL-24，经Pacl线性化后转染QBI-293A包装细胞。收获腺病毒重组病毒子，RT-PCR和Western—blotting鉴定。结果测序结果显示hIL-24序列正确，RT—PCR和Western-blotting检测到了hIL-24的表达。结论成功构建人白介素24的重组腺病毒载体pAdEasy-1-pAdTrack，CMV—hIL-24。获得了人白介素24重组病毒子Ad-hIL-24。     

心衰患者室性心律失常的发生及其预后分析

目的探讨充血性心力衰竭严重程度与室性心律失常的关系及3年内死亡率分析。方法应用24h动态心电图记录分析88例充血性心力衰竭患者(按NYHA心功能分级均在Ⅱ~Ⅳ级),比较各组心律失常发生率并对心功能分级和心律失常程度进行相关性分析。随访44例心力衰竭患者为期3年并比较各组的生存情况。结果心功能Ⅱ级、Ⅲ级和Ⅳ级患者室性心律失常的发生率分别为42.9%、65.7%和88.0%。随着心功能级别的增加,室性心律失常的发生率越高(p=0.003)。心功能分级和心律失常程度之间经Spearman秩相关分析,具有相关性(rs=0.315,p=0.017),心功能分级越高,心律失常程度越严重。3年随访结果显示,各组生存率间有显著性差异(p<0.05),心功能越差,生存率越低。结论充血性心力衰竭越重,室性心律失常发生率越高,程度越严重,其生存率越差。     

婴儿出生后24小时内乙型肝炎疫苗低接种率的原因及居民教育

为防止乙型肝炎 (乙肝 )病毒 (HBV)母婴传播 ,婴儿出生后 2 4h内接种乙肝疫苗 (HepB)非常重要 ,但其接种率很低。为探讨接种率低的原因及其对策 ,在甘肃省访问了承担预防接种的工作人员 ,并对婴儿家长及乡村医生进行了小组访谈。调查结果显示 ,几乎所有婴儿家长对乙肝知识了解甚少或有错误。本次调查中 ,2 4h内接种HepB的婴儿全部在医院出生 ,2 4h内未接种的全部在家出生。即使及时接种的婴儿家长亦不知 2 4h内接种HepB的重要性 ,只不过是因为在医院出生才偶然做到了 2 4h内接种。另外 ,在家分娩且非乡村医生接生时 ,婴儿家长即使知道 2 4h内接种的重要性 ,但如不及时通知乡村医生 ,乡村医生也不能及时到家接种。因此 ,为提高婴儿出生后 2 4h内HepB接种率 ,对婴儿家长的宣传教育 ,特别是对孕妇和新婚夫妇的教育非常必要。     

硫酸镁启动自噬对兔软骨保护作用机制研究

目的探讨硫酸镁通过启动自噬保护兔软骨的作用机制。方法取 24 只成年雌性新西兰兔，采用前交叉韧带切断方法制备创伤性关节炎（post-traumatic osteoarthritis，PTOA）模型，随机分为 PTOA 组、蒸馏水组和硫酸镁组，每组 8 只。术后即刻开始，蒸馏水组及硫酸镁组分别于关节腔内注射 0.5 mL 蒸馏水及 20 mmol/L 硫酸镁溶液，每周 3 次，连续 4 周；PTOA 组不作处理。术后观察动物一般情况；术后 4 周取材，ELISA 检测关节腔积液 TNF-α 和Ⅱ型胶原及静脉血中Ⅱ型胶原表达量，Western blot 检测股骨软骨组织中瞬时感受电位 V5（transient receptor potential channel vanilloid 5，TRPV5）及微管相关蛋白 1 轻链 3（microtubule associated protein 1 light chain 3，LC3）蛋白（LC3-Ⅱ/LC3-Ⅰ）表达，实时荧光定量 PCR 检测滑膜组织中 IL-1β、TNF-α、基质金属蛋白酶 3（matrix metalloproteinases 3，MMP-3）及软骨组织中Ⅱ型胶原、蛋白聚糖（Aggrecan，AGN）、SOX9 mRNA 水平，软骨组织切片行 HE、Masson、阿利新蓝染色并参照改良组织学骨关节炎（osteoarthritis，OA）评分标准评分。结果各组动物均存活至实验完成。与其他两组比较，硫酸镁组关节腔积液中 TNF-α 以及关节腔积液及静脉血中Ⅱ型胶原表达量均降低，TRPV5 蛋白表达明显降低、LC3-Ⅱ/LC3-Ⅰ比值明显升高，滑膜组织中 IL-1β、TNF-α、MMP-3 mRNA 表达降低，软骨组织中Ⅱ型胶原、AGN、SOX9 mRNA 表达升高，OA 评分亦显著降低，差异均有统计学意义（P<0.05）。PTOA 组及蒸馏水组以上指标比较，差异均无统计学意义（P>0.05）。 结论关节腔内注射硫酸镁可减轻兔关节内炎症，减少Ⅱ型胶原及 AGN 丢失，有利于软骨再生，其作用机制可能是通过抑制钙离子通道 TRPV5，进而启动软骨自噬。     

Common cold and influenza symptom management: the use of pharmacokinetic considerations to predict the efficacy of a twice-daily treatment for colds and flu

SUMMARY

Objective: The objective of the two pharmacokinetic studies reported here was to compare the relative bioavailability of an ibuprofen/pseudoephedrine modified-release capsule with each of the active ingredients given alone as standard formulations.

Study design: Evaluation of two open, randomised, cross-over studies, one single dose and one multiple dose, in healthy male volunteers.

Methods: Healthy volunteers were randomised in a cross-over design to single or multiple doses of a combination of ibuprofen (600 mg) plus pseudoephedrine (90 mg) in a slow-release formulation and the individual active products alone as standard formulations; ibuprofen 400mg, pseudoephedrine 60 mg.

Results: The single-dose study demonstrated that the bioavailabilities of ibuprofen and pseudoephedrine achieved with the slow-release formulation were not significantly different from those with standard tablets of each ingredient alone. In addition, mean plasma levels of ibuprofen predictive of clinical efficacy were achieved within 0.5-1 h and lasted for 10-12 h thereafter. The time required to reach clinically effective blood levels of pseudoephedrine was longer, starting at approximately 2 h. However, the plasma levels predicted that the clinical effect would then last for at least a further 12 h. Trough levels from the multiple-dose study showed that clinically relevant analgesic and decongestant plasma levels were maintained for 24 h during twice-daily dosing. The slow-release formulation was well tolerated with only mild adverse events.

Conclusion: Blood levels would predict that the present slow-release fo rmulation of ibuprofen plus pseudoephedrine should offer reliable day and night control of cold and flu and sinus symptoms and be associated with a favourable safety profile.     

Role of asymmetrical dimethylarginine in the progression of renal disease

Asymmetric dimethylarginine (ADMA) is a naturally occurring amino acid found in tissues and cells that circulates in plasma and is excreted in urine. It inhibits nitric oxide synthases (NOs) and produces considerable cardiovascular biological effects. Several studies have suggested that plasma concentrations of ADMA provide a marker of risk for endothelial dysfunction and cardiovascular disease. In animal and in population studies ADMA has been associated with progression of CKD. Several mechanisms may be involved in this association, such as compromise of the integrity of the glomerular filtration barrier and development of renal fibrosis. This review summarizes the existing literature on the biology and physiology of ADMA focusing on its role in the progression of renal disease.     

Steroid Hormone Action in Musculoskeletal Cells Involves Membrane Receptor and Nuclear Receptor Mechanisms

Steroid hormones regulate target cells through traditional nuclear mechanisms as well as by membrane mechanisms. 1 &#102 ,25(OH) 2 D 3 and 24R,25(OH) 2 D 3 bind membrane receptors (mVDR) and mediate their effects on the physiological responses of musculoskeletal cells via protein kinase C (PKC). In cultures of costochondral growth plate chondrocytes, 1 &#102 ,25(OH) 2 D 3 binds the 1,25-mVDR in growth zone cells, activating phospholipase C (PLC), leading to diacylglycerol (DAG) production and PKC translocation to the plasma membrane. It also activates PLA 2 , increasing arachidonic acid release and prostaglandin synthesis. 24R,25(OH) 2 D 3 binds its membrane receptor in resting zone chondrocytes, activating phospholipase D (PLD), and increasing DAG and PKC activity, but translocation does not occur. PLA 2 activity is decreased, reducing arachidonic acid and prostaglandin production. 17 &#103 -Estradiol (E 2 ) activates PKC in both cartilage cells, but DAG is not involved. 1 &#102 ,25(OH) 2 D 3 and 24R,25(OH) 2 D 3 also increase PKC in osteoblasts in a cell-specific manner. Antibodies to the 1,25-mVDR block PKC activation. Membrane-mediated events influence gene expression via signaling cascades, including the ERK1/2 MAP kinases. The ability of steroid hormones to initiate events nongenomically is important for regulation of matrix vesicle (MV) function in the extracellular matrix. MVs have mVDRs, but ligand binding inhibits PKC-zeta (PKC &#145 ) via a mechanism that differs from PKC &#102 activation in the plasma membranes. Treatment of MVs from growth zone chondrocyte cultures with 1 &#102 ,25(OH) 2 D 3 releases stromelysin-1 (MMP-3) and increases TGF- &#103 activation. MMP-3 is also involved in proteoglycan degradation, facilitating calcification. 24R,25(OH) 2 D 3 inhibits PKC &#145 in MV from resting zone cell cultures and inhibits MMP-3 release. Chondrocytes and osteoblasts produce 1,25(OH) 2 D 3 , 24,25(OH) 2 D 3 , and E 2 ; thus, locally produced steroids may function as autocrine regulators of matrix events, including matrix vesicle enzyme activity and matrix protein remodelling during longitudinal growth, calcification, and growth factor activation.     

Estimation of 24-Hour Urinary Sodium Excretion Using Spot Urine Samples

The present study evaluated the reliability of equations using spot urine (SU) samples in the estimation of 24-hour urine sodium excretion (24-HUNa). Equations estimating 24-HUNa from SU samples were derived from first-morning SU of 101 participants (52.4 ± 11.1 years, range 24–70 years). Equations developed by us and other investigators were validated with SU samples from a separate group of participants (n = 224, 51.0 ± 10.9 years, range 24–70 years). Linear, quadratic, and cubic equations were derived from first-morning SU samples because these samples had a sodium/creatinine ratio having the highest correlation coefficient for 24-HUNa/creatinine ratio (r = 0.728, p < 0.001). In the validation group, the estimated 24-HUNa showed significant correlations with measured 24-HUNa values. The estimated 24-HUNa by the linear, quadratic, and cubic equations developed from our study were not significantly different from measured 24-HUNa, while estimated 24-HUNa by previously developed equations were significantly different from measured 24-HUNa values. The limits of agreement between measured and estimated 24-HUNa by six equations exceeded 100 mmol/24-hour in the Bland-Altman analysis. All equations showed a tendency of under- or over-estimation of 24-HUNa, depending on the level of measured 24-HUNa. Estimation of 24-HUNa from single SU by equations as tested in the present study was found to be inadequate for the estimation of an individual’s 24-HUNa.